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Acellular exponential or geometric amplification (e.g., PCR, etc.)

Subclass of:

435 - Chemistry: molecular biology and microbiology

435041000 - MICRO-ORGANISM, TISSUE CELL CULTURE OR ENZYME USING PROCESS TO SYNTHESIZE A DESIRED CHEMICAL COMPOUND OR COMPOSITION

435072000 - Preparing compound containing saccharide radical

435084000 - Preparing nitrogen-containing saccharide

435085000 - N-glycoside

435089000 - Nucleotide

435910100 - Polynucleotide (e.g., nucleic acid, oligonucleotide, etc.)

Patent class list (only not empty are listed)

Deeper subclasses:

Class / Patent application numberDescriptionNumber of patent applications / Date published
435910210 Involving the making of multiple RNA copies 26
Entries
DocumentTitleDate
20130045507Mutagenesis Method - The present invention provides a mutagenesis method wherein a nucleic acid molecule is mutagenized with at least one mutagenesis primer in a primer extension reaction and subsequently amplified by rolling circle amplification (RCA). The method involves steps leading to selective amplification of only the mutated strand by a strand-displacing DNA polymerase. Multiple copies of the mutated plasmids are generated during multiple-primed RCA and the resulting DNA is transformed for use. The method is suitable for mutating both single-stranded and double-stranded DNA. The present invention also provides a kit for use in the mutagenesis method.02-21-2013
20090023189APPARATUS AND METHODS FOR PREPARATION OF SUBTANTIALLY UNIFORM EMULSIONS CONTAINING A PARTICLE - Methods and systems for forming water-in-oil emulsions are described. For example, an apparatus is described which includes: a first compartment containing a plurality of particles dispersed in an aqueous phase; a second compartment containing an oil phase; a porous layer separating the first and second compartments; and a device for applying pressure to the first compartment. A method is described which includes: moving an oil phase relative to a surface of a porous layer while simultaneously forcing an aqueous composition comprising particles through the porous layer and into the flowing dispersion medium thereby forming droplets of the aqueous composition containing particles dispersed in the oil phase. The aqueous composition can include one or more nucleic acid templates and reagents for amplifying the nucleic acids such as PCR reagents. A porous partition is described comprising a first and second major surfaces and at least two straight through pores comprising a cross sectional shape selected from a polygon, an oval, an oblong, a dumbbell, a bowtie and irregular shapes thereof. Aqueous droplets containing an oligonucleotide attached to a particle and reagents can be used as a microreactor for nucleic acid amplification.01-22-2009
20080261276Micro-Fluidic Device Based Upon Active Matrix Principles - A micro-fluidic device (10-23-2008
20120183998Modified Promoter - The present invention provides a modified promoter, an expression vector and a transformant each containing the promoter, and a method for producing a gene product of interest using the transformant. The invention provides a modified promoter, including a nucleotide sequence of a promoter derived from bacterium belonging to the genus 07-19-2012
20100151531METHOD FOR AMPLIFICATION OF DNA FROM BLOOD SAMPLE AND DNA AMPLIFICATION KIT THEREFOR - A DNA amplification method including: subjecting a blood sample having DNA to be amplified, to a pretreatment using an alkaline aqueous solution under ordinary temperature, so as to extract double-stranded DNA from the blood sample and dissociate the double-stranded DNA into a single-stranded DNA to obtain a blood-derived sample including the single-stranded DNA; preparing an isothermal amplification reaction solution comprising a mixture of the blood-derived sample, a primer, dNTP, a strand-displacing DNA polymerase, a magnesium salt and a buffer, to establish an isothermal amplification reaction system meeting optimum conditions for the strand-displacing DNA polymerase; and amplifying DNA in the isothermal amplification reaction system using the single-stranded DNA as a template.06-17-2010
20110195458Method and Apparatus for Amplifying Nucleic Acid Sequences - A method of amplifying one or more target nucleic acid sequence(s) in a PCR buffer solution comprising one or more target nucleic acid sequence(s), a nucleic acid polymerase, deoxynucleotide or nucleotide triphosphates, at least a first primer and a second primer is provided. The PCR buffer solution is contained in an elongated vessel. The solution within the vessel is locally heated in a spatio-temporally varying manner. Fluid flows circulating within the vessel are established. At least a fraction of the PCR buffer solution cyclically travels between regions having different temperatures, giving rise to a temperature cycling.08-11-2011
20110195457ISOTHERMAL AMPLIFICATION OF NUCLEIC ACID USING PRIMERS COMPRISING A RANDOMIZED SEQUENCE AND SPECIFIC PRIMERS AND USES THEREOF - Methods and kits for amplifying a nucleic acid under isothermal conditions to form an amplified nucleic acid sequence are provided. The methods and kits comprises providing a nucleic acid template, a DNA polymerase, deoxyribonucleoside triphosphates, a primer comprising a randomized sequence, and a specific primer, and amplifying the nucleic acid template.08-11-2011
20110195459Methods of Making Libraries of Nucleic Acids Using Porous Particles - The invention provides particle compositions having applications in nucleic acid analysis. Nucleic acid polymer particles of the invention allow polynucleotides to be attached throughout their volumes for higher loading capacities than those achievable solely with surface attachment. In one aspect, nucleic acid polymer particles of the invention comprise polyacrylamide particles with uniform size distributions having low coefficients of variations, which result in reduced particle-to-particle variation in analytical assays. Such particle compositions are used in various amplification reactions to make amplicon libraries from nucleic acid fragment libraries.08-11-2011
20100081174Methods for Detection of Methyl-CpG Dinucleotides - The invention provides methods for enriching methyl-CpG sequences from a DNA sample. The method makes use of conversion of cytosine residues to uracil under conditions in which methyl-cytosine residues are preserved. Additional methods of the invention enable to preservation of the context of me-CpG dinucleotides. The invention also provides a recombinant, full length and substantially pure McrA protein (rMcrA) for binding and isolation of DNA fragments containing the sequence 5′-C04-01-2010
20100120099Device For The Carrying Out Of Chemical or Biological Reactions - The invention relates to a device for the carrying out of chemical or biological reactions with a reaction vessel receiving element for receiving a microtiter plate with several reaction vessels, wherein the reaction vessel receiving element has several recesses arranged in a regular pattern to receive the respective reaction vessels, a heating device for heating the reaction vessel receiving element, and a cooling device for cooling the reaction vessel receiving element.05-13-2010
20100120100Device For The Carrying Out of Chemical or Biological Reactions - An interactive radio network enables users to interact with the content of a radio broadcast, including commercials or messages, and to selectively save, store, review, fast forward, rewind, pause, forward, and respond to the radio programs and/or the commercials. The interactive radio network provides a widespread, international, and economical access to the radio stations, and reduces the need for advertisement billboards. It provides the users with an opportunity to selectively inquire about the products or services being advertised. Furthermore, the interactive radio network allows the users as well as various sectors of the advertisement industry to interact with the content of the radio broadcast. The advertisements are no longer limited to audio messages, but can further include elaborate video, text, and data information. The interactive radio network enables the users to communicate and interact with each others, based on the broadcast content. It also provides a widely accessible and affordable avenue for mass marketing and broadcasting of commercials to mobile users.05-13-2010
20130078676METHOD OF REGULATING OLIGONUCLEOTIDE FUNCTIONALITY - The present invention relates generally to a method of regulating oligonucleotide functionality and, more particularly, to a method of regulating the functionality of a primer or probe. The method of the invention is designed to provide a means to selectively inactivate or activate the functionality of an oligonucleotide, such as a primer, thereby providing means to regulate the progress of any method using that oligonucleotide. The development of a means to regulate the functionality of an oligonucleotide, such as a primer, is useful in a range of applications including, but not limited to, amplification reactions such as PCR, isothermal amplification and nucleic acid strand extension. With respect to amplification reactions, these have wide utility including the diagnosis and/or monitoring of disease conditions which are characterised by specific gene sequences and the characterisation or analysis of specific gene regions of interest.03-28-2013
20100075384Helicase-dependent amplification of circular nucleic acids - A helicase-mediated amplification method for circular DNA templates and target DNA sequences within the templates is provided. The method combines a DNA polymerase and a helicase preparation to amplify a target sequence as well as the entire circular DNA template.03-25-2010
20100159527Polypeptides Having Nucleic Acid Binding Activity and Compositions and Methods For Nucleic Acid Amplification - Polypeptides having nucleic acid binding activity are provided. Methods of using polypeptides having nucleic acid binding activity are provided. Fusion proteins and methods of using fusion proteins are provided. Fusion proteins comprising a polymerase and a nucleic acid binding polypeptide are provided. Fusion proteins comprising a reverse transcriptase and a nucleic acid binding polypeptide are provided. Methods are provided for amplifying a nucleic acid sequence using a fusion protein comprising a nucleic acid binding polypeptide and a polymerase. Methods are provided for amplifying a nucleic acid sequence using a fusion protein comprising a nucleic acid binding polypeptide and a reverse transcriptase.06-24-2010
20130084606Composition for Preventing Evaporation of Reaction Solution During Nucleic Acid Amplification Reaction - The present invention provides a composition capable of hermetically sealing a PCR reaction vessel without using a closure member or adhesive seal. Disclosed is a composition for preventing evaporation of a nucleic acid amplification reaction solution during nucleic acid amplification reaction, which is a liquid during the reaction and becomes a solid through chemical or thermal changes after completion of the reaction. Also disclosed is a composition for preventing evaporation of a nucleic acid amplification reaction solution during nucleic acid amplification reaction, wherein the melting point of the composition is 0-15° C. Also disclosed is a composition for preventing evaporation of a nucleic acid amplification reaction solution during nucleic acid amplification reaction, wherein the melting point of the composition is 5-10° C.04-04-2013
20130034879DNA Polymerases - A DNA polymerase mutant comprising a Taq DNA polymerase amino acid sequence with a mutation at one or more of the following selected amino acid positions: E189K, E230K, E507K, H28R, L30R, G38R, F73V, H75R, E76A, E76G, E76K, E90K, K206R, E315K, A348V, L351F, A439T, D452N, G504S, E507A, D551N, L552R, I553V, D578N, H676R, Q680R, D732G, E734G, E734K, F749V; wherein the polymerase mutant exhibits relative to wild-type DNA polymerase increased polymerase speed, increased affinity to DNA substrate and/or increased resistance to a DNA polymerase inhibitor; and wherein, when the mutation is E507K in combination with two or more further mutations or the mutation is Q680R in combination with four or more further mutations, at least one of the further mutations is at one of the selected amino acid positions; and when the mutation is I553V, this is not in combination with D551S.02-07-2013
20130040343Methods for Detection of Methyl-CpG Dinucleotides - The invention provides methods for enriching methyl-CpG sequences from a DNA sample. The method makes use of conversion of cytosine residues to uracil under conditions in which methyl-cytosine residues are preserved. Additional methods of the invention enable to preservation of the context of me-CpG dinucleotides. The invention also provides a recombinant, full length and substantially pure McrA protein (rMcrA) for binding and isolation of DNA fragments containing the sequence 5′-C02-14-2013
20100003723RNA-DEPENDENT DNA POLYMERASE FROM GEOBACILLUS STEAROTHERMOPHILUS - The invention relates to an isolated polynucleotide sequence from the genome of 01-07-2010
20090155856NUCLEIC ACID AMPLIFICATION METHOD - An object to be achieved by the present invention is to provide a nucleic acid amplification method by which a nucleic acid can be amplified using oligonucleotide primers and DNA polymerase. The present invention provides a nucleic acid amplification method which comprises performing incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase having strand displacement activity, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3′ end of the primer and thus amplifying the nucleic acid fragment, wherein a first oligonucleotide primer and a second oligonucleotide primer are designed in such a way that a region which contains two identical sequences X of serial 4 or more nucleotides within, the region of 200 or less nucleotides, or apart thereof can be amplified.06-18-2009
20090155855COVER FOR SAMPLE WITH HOMOGENOUS PRESSURE APPLICATION - The present invention relates to means for covering one or more sample(s) that are suitable to avoid or minimize evaporation and/or condensation of any vaporizable substance that may be present in the sample(s) or reaction mixture(s), in particular evaporation of substance at the fringes of a vessel or an array of vessels or condensation of said substance on the lid of a reaction vessel or a plate/block containing the sample(s) and/or the means for covering. This is achieved by providing a device comprising, among others, a force distribution unit that comprises at least one medium or material that is unable to withstand a static shear stress and deforms continuously under the action of a shear force. In a preferred embodiment, this medium or material is a gas, a shear force.06-18-2009
20090155854METHOD FOR AMPLIFYING A FLAVIVIRUS cDNA IN A PROKARYOTIC CELL - The invention relates to a method for amplifying a functional flavivirus cDNA in a prokaryotic cell, such as 06-18-2009
20100105109MULTIPLY-PRIMED AMPLIFICATION OF CIRCULAR NUCLEIC ACID SEQUENCES - Improved processes for the amplification of target DNA sequences in the form of single or double stranded circular DNA molecules, especially those present in colony and plaque extracts, using multiple specific and/or random sequence oligonucleotide primers are disclosed. The product of this amplification is used for analysis by restriction enzyme digestion or DNA sequencing and other analyses that involve hybridization. Kits containing components for use in the method are also described. Also described are further uses of this amplified DNA in sequencing, genotyping and haplotyping, and other molecular biology applications.04-29-2010
20100041105DNA IN THE PRESENCE OF GELLAN - A method is provided for nucleic acid amplification with enhanced sensitivity. The method for enhanced sensitivity involves carrying out the amplification reaction in the presence of gellan. For instance, the method allows for the production of detectable amounts of PCR amplified DNA from at least 10 fold fewer target molecules than a comparable PCR reaction in absence of gellan.02-18-2010
20080199915Methods and Kits For Mass Production Of Dsrna - The invention relates to methods and kits for propagating target nucleic acid in the form of double stranded RNA. This invention relates in particular to a method for mass production of dsRNA. The method comprises that a target nucleic acid is provided in a form replicable by an RNA-dependent RNA polymerase in a living cell, said replicable form of the target nucleic acid is contacted with said polymerase under conditions sufficient for template-directed RNA synthesis, wherein one of the reaction products is necessarily double-stranded (ds) RNA and said dsRNA products are recovered in a sufficiently pure form. The dsRNA products can be used in various applications, for example in gene silencing.08-21-2008
20100330619DIRECT MULTIPLEX CHARACTERIZATION OF GENOMIC DNA - The invention is directed to novel methods of multiplexing nucleic acid reactions, including amplification, detection and genotyping. The invention relies on the use of precircle probes that are circularized in the presence of the corresponding target nucleic acids, cleaved, and then amplified.12-30-2010
20120164691COMPOSITIONS AND METHODS FOR PRODUCING SINGLE-STRANDED CIRCULAR DNA - The present invention provides methods, kits, and compositions for producing single-stranded circular DNA by PCR. In particular, hairpin primers are provided, and methods of use thereof to produce single-stranded circular DNA molecules.06-28-2012
20090123976Compositions and Method for Storage of Nucleic Acid From Bodily Fluids - An aqueous composition comprising a denaturing agent, a chelator, a buffering agent and a protease for the extraction of nucleic acid from a sample of bodily fluid, such as saliva, such that the extracted nucleic acid is stable for at least fourteen days at room temperature and can be directly utilised in an amplification reaction without further processing. In particular, said composition comprises SDS, Cyclohexanediamine tetraacetate, Tris-HCl and proteinase K. A method and kit for the amplification of DNA directly from a bodily fluid, comprising said composition, is further provided.05-14-2009
20090325238Method of Analyzing a BRCA2 Gene in a Human Subject - Five novel DNA and protein sequences have been determined for the BRCA2 gene, as have been ten polymorphic sites and their rates of occurrence in the normal alleles of BRCA2. The sequences BRCA212-31-2009
20130089895Polypeptides Having Nucleic Acid Binding Activity and Compositions and Methods For Nucleic Acid Amplification - Polypeptides having nucleic acid binding activity are provided. Methods of using polypeptides having nucleic acid binding activity are provided. Fusion proteins and methods of using fusion proteins are provided. Fusion proteins comprising a polymerase and a nucleic acid binding polypeptide are provided. Fusion proteins comprising a reverse transcriptase and a nucleic acid binding polypeptide are provided. Methods are provided for amplifying a nucleic acid sequence using a fusion protein comprising a nucleic acid binding polypeptide and a polymerase. Methods are provided for amplifying a nucleic acid sequence using a fusion protein comprising a nucleic acid binding polypeptide and a reverse transcriptase.04-11-2013
20090311754Polynucleotide Amplification - The present invention provides a method for amplifying a pool of polynucleotide molecules in a sample, characterized by the steps of a) obtaining a sample or RNA and reverse transcription of entire RNA molecules thus creating full length cDNA or obtaining a sample of full length cDNA, b) tailing the 3′ end of the transcribed cDNA with a polynucleotide tail after the 3′ end, c) amplification of the cDNA using a pair of primers, wherein a first 3′ primer is specific for the 5′ end of the cDNA and a second 5′ primer is specific for the a upstream portion of the polynucleotide tail and the next 1 to 10 nucleotides upstream of the 3′polynucleotide tail of the cDNA.12-17-2009
20130071879AMPLIFICATION METHODS TO MINIMISE SEQUENCE SPECIFIC BIAS - Methods for amplifying nucleic acids are provided. The methods can be used to minimise sequence specific bias caused by the preferential amplification of certain nucleic acid sequences. Methods are described which can lower the efficiency of AT rich templates relative to GC rich templates, thereby minimising GC bias during amplification reactions with multiple templates of different sequence. The methods are suited to solid phase amplification, for example, utilising flow cells.03-21-2013
20130071880Dna amplification apparatus and method - The invention relates to a method for the amplification of DNA and apparatus for such amplification. The method comprises steps typically found in amplification methods but utilises an optical procedure to detect denaturation of DNA or attain03-21-2013
20090280538Methods and compositions for nucleic acid sample preparation - Provided are methods and compositions for the production of linear single-stranded nucleic acids, which can be used as templates in high-throughput sequencing systems. Also provided are methods and compositions for the production of closed single-stranded nucleic acid loops, which can be used as templates in high-throughput sequencing systems.11-12-2009
20130164788COMPOSITIONS AND METHODS FOR REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION (RT-PCR) - The present invention is directed to compositions and methods useful for the amplification of nucleic acid molecules by reverse transcriptase-polymerase chain reaction (RT-PCR). Specifically, the invention provides compositions and methods for the amplification of nucleic acid molecules in a simplified one- or two-step RT-PCR procedure using combinations of reverse transcriptase and thermostable DNA polymerase enzymes in conjunction with sulfur-containing molecules or acetate-containing molecules (or combinations of such sulfur-containing molecules and acetate-containing molecules), and optionally bovine serum albumin. The invention thus facilitates the rapid and efficient amplification of nucleic acid molecules and the detection and quantitation of RNA molecules. The invention also is useful in the rapid production and amplification of cDNAs which may be used for a variety of industrial, medical and forensic purposes.06-27-2013
20110014660THERMOSTABLE DNA POLYMERASE FROM PALAEOCOCCUS FERROPHILUS - There is provided a polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 90% identity to 01-20-2011
20110014659ISOLATION OF UNKNOWN REARRANGED T-CELL RECEPTORS FROM SINGLE CELLS - Disclosed herein are methods and materials for isolating and identifying T cell receptors from single cells. In some embodiments, genomic DNA from a single T cell is isolated using whole genome amplification (WGA). A series of PCR reactions is carried out to enrich the genomic template for sequences encoding the TCR alpha and beta chains, and then to isolate the sequences encoding the TCR alpha and beta chains.01-20-2011
20110014658COMPOSITIONS AND METHOD FOR STORAGE OF NUCLEIC ACID FROM BODILY FLUIDS - The present invention provides an aqueous composition and method for extracting nucleic acid from a sample of bodily fluid, such as saliva, such that the nucleic acid within said sample remains stable for at least fourteen days at room temperature. The composition permits direct use of the extracted and stored DNA in an amplification reaction without further processing.01-20-2011
20130059342COMPOSITIONS AND METHODS FOR USE IN RECOMBINATIONAL CLONING OF NUCELIC ACIDS - The present invention relates to compositions and methods for use in recombinational cloning of nucleic acid molecules. In particular, the invention relates to nucleic acid molecules encoding one or more recombination sites or portions thereof, to nucleic acid molecules comprising one or more of these recombination site nucleotide sequences and optionally comprising one or more additional physical or functional nucleotide sequences. The invention also relates to vectors comprising nucleic acid molecules of the invention, to host cells comprising vectors or nucleic acid molecules of the invention, to methods of producing polypeptides using nucleic acid molecules of the invention, and to polypeptides encoded by these nucleic acid molecules or produced by methods of the invention. The invention also relates to the use of these compositions in methods for recombinational cloning of nucleic acids, in vitro and in vivo, to provide chimeric DNA molecules that have particular characteristics and/or DNA segments.03-07-2013
20120190074THERMOCYCLER AND SAMPLE PORT - The invention relates to continuous flow systems, in particular thermocyclers for the automated and continuous cycling of fluid between a plurality of temperature zones in the amplification of nucleic acids. The invention also relates to an improved sample port for introducing a volume of a liquid sample into a continuous flow system.07-26-2012
20090130719Microfluidic Cartridge - The technology described herein generally relates to microfluidic cartridges configured to amplify and detect polynucleotides extracted from multiple biological samples in parallel. The technology includes a microfluidic substrate, comprising: a plurality of sample lanes, wherein each of the plurality of sample lanes comprises a microfluidic network having, in fluid communication with one another: an inlet; a first valve and a second valve; a first channel leading from the inlet, via the first valve, to a reaction chamber; and a second channel leading from the reaction chamber, via the second valve, to a vent.05-21-2009
20090269815LINEAR AMPLIFICATION OF SHORT NUCLEIC ACIDS - The present teachings provide novel methods for amplifying short nucleic acids. In some embodiments, the present teachings provide novel methods for linearly amplifying a collection of micro RNAs by using temperature cycling during a reverse transcription reaction. The cycling can comprise at least 20 cycles of an annealing temperature segment of 10° C.-30° C., and a denaturation temperature segment of 35° C.-60° C. In some embodiments, the temperature cycled reaction can comprise an osmolyte.10-29-2009
20090269814Method of Analyzing a BRCA2 Gene in a Human Subject - Five novel DNA and protein sequences have been determined for the BRCA2 gene, as have been ten polymorphic sites and their rates of occurrence in the normal alleles of BRCA2. The sequences BRCA210-29-2009
20090269813Methods For Multiplexing Recombinase Polymerase Amphlification - This disclosure provides for methods and reagents for rapid multiplex RPA reactions and improved methods for detection of multiplex RPA reaction products. In addition, the disclosure provides new methods for eliminating carryover contamination between RPA processes.10-29-2009
20120225456METHOD FOR CONDUCTING MULTIPLE REACTIONS IN A SINGLE REACTION TUBE - There is disclosed a method for conducting at least two reactions in a reaction tube, said method comprising the steps of providing at least two reaction phases within said reaction tube for allowing said reactions to occur therein, providing a separation phase that is immiscible with said two reaction phases and which is disposed therebetween, providing at least one particle capable of being coupled to a chemical species, wherein said particle is movable between said reaction phases to introduce said chemical species thereto.09-06-2012
20130065281METHOD FOR SYNTHESIZING cDNA - A method for synthesizing cDNA characterized by preparing a reaction solution that does not allow an endodeoxyribonuclease to show its activity, without thermal deactivation of the endodeoxyribonuclease or removal of the endodeoxyribonuclease, and carrying out a reverse transcription reaction, wherein the reaction solution contains a treated sample and a reverse transcriptase, the treated sample being formed by treating a sample comprising RNA and DNA with the endodeoxyribonuclease to degrade DNA in the sample. The method and the kit for synthesizing cDNA of the present invention are widely useful in genetic engineering fields.03-14-2013
20130065280MICROFLUIDIC APPARATUS AND CONTROL METHOD THEREOF - A microfluidic apparatus having an additional chamber containing material configured to prevent cross contamination between reaction chambers contained therein, and a control method thereof are provided. The microfluidic apparatus includes a sample chamber configured to accommodate a sample, a plurality of reaction chambers each configured to accommodate a reagent, a distribution channel configured to distribute the sample into the plurality of reaction chambers, a mixture prevention chamber connected to the distribution channel and containing a mixture prevention material configured to prevent the reagents accommodated in the plurality of reaction chambers from being mixed with each other, and a valve disposed within the distribution channel and configured to open and close the distribution channel.03-14-2013
20130164789System and Methods for Making and Processing Emulsions - An automated template bead preparation system is provided and includes a membrane-based emulsion generation subsystems, a thermal plate and subsystem, and a continuous centrifugation emulsion breaking and templated bead collection subsystem. The emulsion generation subsystem provides uniformity in the preparation of an inverse emulsion and may be used to create large or small volume inverse emulsions rapidly and reproducibly. An emulsion-generating device is provided that can supply a continuous stream of an inverse emulsion to a thermal subsystem, in automated fashion. The thermal subsystem can treat an inverse emulsion passed therethrough. The continuous centrifugation subsystem can continuously break a thermally cycled inverse emulsion and collect template beads formed in the aqueous microreactor droplets of the inverse emulsion.06-27-2013
20110020879REACTION DEVICE, REACTION METHOD AND METHOD OF SYNTHESIZING cDNA - The present invention provides a device and a method whereby plural kinds of reaction operations and washing operations can be conducted in parallel without washing or replacing an instrument used in transferring a solution and the like in each operation. A reaction device having a plurality of projecting barriers provided in a line on one surface of a substrate, wherein the projecting barrier has at least one cutoff portion and an inner space capable of holding a droplet, and at least the face holding the droplets of the substrate surface has a contact angle to pure water of from 90 to 150 degrees. A reaction method using the above reaction device wherein a substance immobilized on magnetic beads is sequentially transferred in and between droplets of a solution containing a surface tension reducing agent that are held in the spaces for holding a droplet defined by the above-described projecting barriers by means of a magnet located on the opposite surface of the substrate to the surface having the projecting barriers to thereby conduct reactions and washings.01-27-2011
20110020878Method of Removing Nucleic Acid Contamination in Reverse Transcription and Amplification Reactions - The invention provides methods of removing nucleic acid contamination from reverse transcription reactions and hot-start PCR, wherein said hot-start PCR is a barrier hot-start PCR set up and/or involves a hot-start DNA polymerase, which methods comprise use of a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA. The invention further provides a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA, nucleic acids encoding said DNase and kits or compositions comprising said DNase or said nucleic acid.01-27-2011
20110020877CREN7 CHIMERIC PROTEIN - There is provided a chimeric protein comprising a nucleic acid modifying enzyme domain having nucleic acid modifying activity joined with an Cren7 enhancer domain or variant thereof, in which the Cren7 enhancer domain or variant thereof enhances the activity of the nucleic acid modifying enzyme domain compared with a corresponding protein lacking the Cren7 enhancer domain or variant thereof. There is also provided an isolated nucleic acid encoding the chimeric protein of the invention and methods utilising the protein.01-27-2011
20110020876MESOSCALE POLYNUCLEOTIDE AMPLIFICATION DEVICES - Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide amplification reaction. The devices are provided with a substrate microfabricated to include a polynucleotide amplification reaction, chamber, having at least one cross-sectional dimension of about 0.1 to 1000 μm. The device also includes at least one port in fluid communication with the reaction chamber, for introducing a sample to the chamber, for venting the chamber when necessary, and, optionally, for removing products or waste material from the device. The reaction chamber may be provided with reagents required for amplification of a preselected polynucleotide. The device also may include means for thermally regulating the contents of the reaction chamber, to amplify a preselected polynucleotide. Preferably, the reaction chamber is fabricated with a high surface to volume ratio, to facilitate thermal regulation.01-27-2011
20090047713Microfluidic Cartridge and Method of Making Same - The present technology provides for a microfluidic substrate configured to carry out PCR on a number of polynucleotide-containing samples in parallel. The substrate can be a single-layer substrate in a microfluidic cartridge. Also provided are a method of making a microfluidic cartridge comprising such a substrate. Still further disclosed are a microfluidic valve suitable for use in isolating a PCR chamber in a microfluidic substrate, and a method of making such a valve.02-19-2009
20090047712Chemically cleavable phosphoramidite linkers - The present invention provides phosphoramidite linkers that are useful for the production of synthesizing two or more oligonucleotides in tandem. The inventive linkers have the following desirable properties: (i) enhanced stability to alkali conditions versus the linkers previously published, (ii) cleave to produce 5′ and 3′ ends that are fully biologically compatible, (iii) cleave completely under conditions that are already used in cleavage/deprotection processes so they are fully compatible with conditions that are common in laboratories and do not require additives that necessitate further purification after cleavage, (iv) integrate easily onto commercially available synthesizers because they are compatible with standard coupling chemistry, and (v) are compatible with DNA, RNA, forward, reverse, and synthesis chemistries. In addition, the inventive linkers may be coupled to a solid support. Thus, the inventive linkers provide a significant advancement in the state of the art.02-19-2009
20120115190AIR COOLING SYSTEMS AND METHODS FOR MICROFLUIDIC DEVICES - Systems and methods for air cooling a microfluidic device using confinement channels to isolate cooling air from exposed liquids are disclosed. The systems and methods may also thermally condition the cooling airflow for improved robustness of the microfluidic device. In one embodiment, the air cooling system includes a split-level cooling manifold including an inlet duct that directs cooling air to a microfluidic device and an outlet duct that directs air heated by the microfluidic device away from the microfluidic device. The temperature of cooling air may be measured. The cooling air may be preheated to a temperature that is higher than an expected ambient temperature. The temperature of the cooling air after being heated by a microfluidic device may be measured.05-10-2012
20120115189MICROFLUIDIC AND NANOFLUIDIC DEVICES, SYSTEMS, AND APPLICATIONS - The present invention discloses the integration of programmable microfluidic circuits to achieve practical applications to process biochemical and chemical reactions and to integrate these reactions. In some embodiments workflows for biochemical reactions or chemical workflows are combined. Microvalves such as programmable microfluidic circuit with Y valves and flow through valves are disclosed. In some embodiments microvalves of the present invention are used for mixing fluids, which may be part of an integrated process. These processes include mixing samples and moving reactions to an edge or reservoir for modular microfluidics, use of capture regions, and injection into analytical devices on separate devices. In some embodiments star and nested star designs, or bead capture by change of cross sectional area of a channel in a microvalve are used. Movement of samples between temperature zones are further disclosed using fixed temperature and movement of the samples by micropumps.05-10-2012
20090029422REACTOR PLATE AND REACTION PROCESSING METHOD - The reactor plate preferably includes a sealed reactor, a reactor flow channel connected to the reactor, a sample container constituted from a sealed container provided separately from the reactor, a sample container flow channel to be connected to the sample container, a syringe for sending a liquid, a switching valve for connecting the syringe to the reactor flow channel or the sample container flow channel, and a projecting flow channel connected to the end of the sample container flow channel located on the sample container side. The sample container has a penetrable portion through which the projecting flow channel can penetrate and which is provided to be opposed to the projecting flow channel and is located at a position such that the projecting flow channel penetrating the penetrable portion is brought into contact with a liquid contained in the sample container. The sample container can be connected to the sample container flow channel by inserting the tip of the projecting flow channel into the sample container through the penetrable portion.01-29-2009
20090029421Recombinase polymerase amplification - The present invention features novel, diverse, hybrid and engineered recombinase enzymes, and the utility of such proteins with associated recombination factors for carrying out DNA amplification assays. The present invention also features different recombinase ‘systems’ having distinct biochemical activities in DNA amplification assays, and differing requirements for loading factors, single-stranded DNA binding proteins (SSBs), and the quantity of crowding agent employed.01-29-2009
20100086975METHOD AND APPARATUS FOR AMPLIFICATION OF NUCLEIC ACID SEQUENCES USING IMMOBILIZED DNA POLYMERASE - The present invention generally relates to methods and apparatuses for amplifying nucleic acid sequences using immobilized DNA polymerase. More particularly, it relates to methods and apparatuses useful for amplifying target nucleic acid sequences by forming a plurality of reaction regions in which polymerase chain reaction (PCR) can occur, positioning immobilized DNA polymerase in a specific reaction region, and circulating DNA through the reaction regions. The present invention provides those methods and apparatuses that allow simple separation and recovery of the DNA polymerase after the amplification, that can be operated not only with thermostable DNA polymerases but also with non-thermostable DNA polymerases, and that are simpler in their designs and processes so that they can be readily integrated into complex devices such as Lab-on-a-chip.04-08-2010
20100086976METHOD FOR DECONTAMINATING A SOLUTION WITH RESPECT TO UNWANTED NUCLEIC ACIDS - The present invention concerns a method for decontaminating a solution of any nucleic acid present in said solution, comprising the following steps: 04-08-2010
20100086977Pressure Chamber Clamp Mechanism - In a high-density sequence detection system, an apparatus for transporting a microplate. A control system provides a thermocycler control signal to regulate a desired thermal output of a thermocycler block. A frame has a first side, a second side, and an opening that extends through the first side and the second side. A window is positioned in the opening and a circumferential seal is positioned around a periphery of the opening at the second side. The seal has a peripheral lip that seals against the microplate. A clamp is adapted to engage and displace the frame from an unclamped position to a clamped position, wherein in the unclamped position, the microplate is displaced away from the thermocycler block and in the clamped position, the clamp urges the microplate against the thermocycler block. In some embodiments the peripheral lip is positioned radially inward of the periphery of the opening.04-08-2010
20080268508Methods and kits for negative selection of desired nucleic acid sequences - The present invention pertains to a method to isolate, separate, enrich or amplify a targeted nucleotide polymer such as mRNA through selective reverse transcription of the targeted polymer into cDNA from a sample comprising of chemically identical or similar polynucleotide polymers such as rRNA. The enrichment of the targeted nucleic acid such as mRNA is accomplished by blocking the reverse transcription of undesired rRNA while allowing unrestricted reverse transcription of the targeted polymer. The invention also embodies that the cleavage of the non-targeted nucleic acid such as rRNA bound to an oligonucleotide through enzymatic activity (RNase H). The invention further embodies methods and kits to accomplish the utility of the invention through the following steps 1) 3′ tailing of chemically identical or similar nucleotide polymers in a sample that includes bacterial mRNA 2) a 3′ tail capable of binding to a oligo-dN primer 3) at least one oligonucleotide capable of preventing the extension of oligo-dN bound to at least one non-targeted nucleotide polymers by a DNA polymerase such as a reverse transcriptase without restricting conversion of bacterial mRNA into cDNA 4) where the non-targeted molecule is prevented as a template for cDNA synthesis by enzymatic cleavage (RNase H) of template (rRNA)-oligonucleotide hybrid 5) where the reverse transcriptase is physically blocked by the oligonucleotide bound to the non-targeted nucleic acids such as rRNA 5) purification of the selectively transcribed cDNA. In further embodiments of the present invention, methods and composition to enable the study of bacterial transcriptomics-an analysis of genes expressed by a bacterial infection of a host, an isolated bacterial culture or a bacterial community, such as recovered from soil, intestine, mouth, biofilm, water etc are also included for use in DNA-chip or sequencing analyses.10-30-2008
20120270272DETECTION OF NUCLEIC ACID SEQUENCE DIFFERENCES USING COUPLED LIGASE DETECTION AND POLYMERASE CHAIN REACTIONS - The present invention relates to a method for identifying a target nucleotide sequence. This method involves forming a ligation product on a target nucleotide sequence in a ligation detection reaction mixture, amplifying the ligation product to form an amplified ligation product in a polymerase chain reaction (PCR) mixture, detecting the amplified ligation product, and identifying the target nucleotide sequence. Such coupling of the ligase detection reaction and the polymerase chain reaction permits multiplex detection of nucleic acid sequence difference.10-25-2012
20090130720METHODS AND KITS FOR REDUCING NON-SPECIFIC NUCLEIC ACID AMPLIFICATION - Methods and kits for efficient amplification of nucleic acids are provided. The methods comprise in-vitro amplification of a nucleic acid template employing partially constrained primers having terminal mismatch primer-dimer structure. The methods also comprise in-vitro amplification of a nucleic acid template employing partially constrained primers having nucleotide analogues. The methods enhance efficiency of nucleic acid amplification reaction by reducing non-specific amplification reactions.05-21-2009
20120045799THERMOCYCLER SEAL COMPOSITION, METHOD, AND APPLICATION - An immiscible mixture of wax and silicone oil is provided for application to the inner surface of a thermocycle reactor tube. The mixture is a solid at typical room temperatures and under typical product storage conditions, but at temperatures above the melting point of the mixture (e.g., >46° C. and therefore, at or above 90° C. (which is typical of the first stage of a PCR thermocycle)) the mixture melts and covers the exposed surface of the solution undergoing the thermocycle, thereby sealing the reaction against evaporation and/or condensation over the duration of the multiple thermocycles typical of a PCR reaction. Upon cooling of the reaction tube, the resulting amplicons can then extracted from below the sealing layer.02-23-2012
20100261229SYSTEM AND METHOD FOR PREPARING AND USING BULK EMULSION - An emulsion generation apparatus and method for forming an emulsion are provided wherein a customized impeller design is adapted to form an emulsion with a desired droplet size that defines a desired volume. The emulsion generation apparatus provides improved uniformity in emulsion preparation and may be used to create large or small volume emulsions rapidly and reproducibly. A system and method are also provided for large volume sample amplification adaptable for use with conventional PCR-based reactions as well as emulsion-based PCR reactions and other reactions. For applications involving emulsion-based PCR amplification, the system and method provide improved uniformity in emulsion amplification and can be used to amplify large or small volume emulsions rapidly and reproducibly.10-14-2010
20110281305MODIFIED TYPE A DNA POLYMERASES - The present invention provides improved DNA polymerases, in particular, type A DNA polymerases, that may be better suited for applications in recombinant DNA technologies. Among other things, the present invention provides modified DNA polymerases derived from directed evolution experiments designed to select mutations that confer advantageous phenotypes under conditions used in industrial or research applications.11-17-2011
20090137008REDUCED INHIBITION OF ONE-STEP RT-PCR - The present invention provides a method for amplifying a nucleic acid molecule. The method involves mixing an RNA template with a composition having a reverse transcriptase, a DNA polymerase and a RT inhibition reducer. The RT inhibition reducer can be Sso7d, Sac7d, Sac7e, Sso7e, AluI methylase, suramin, a phosphorothioate oligodeoxycytosine, a phosphorothioate oligodeoxyadenine, a phosphorothioate oligodeoxythymine or poly(rA)(dT). The mixing forms a mixture that is incubated under conditions sufficient to synthesize a DNA molecule complementary to all or a portion of the RNA template, thereby amplifying the nucleic acid molecule.05-28-2009
20090148912BIOLOGICAL SAMPLE REACTION CHIP, BIOLOGICAL SAMPLE REACTION APPARATUS, AND BIOLOGICAL SAMPLE REACTION METHOD - A biological sample reaction chip, including: a plurality of reactors disposed on one plane; a reaction fluid distribution channel connected via a microchannel to each reactor and provided on the plane on which the plurality of reactors are disposed; and a reaction fluid movement stopping unit, which is connected to an end point of the reaction fluid distribution channel and is capable of controlling movement of a reaction fluid.06-11-2009
20090291475SEQUENCE AMPLIFICATION WITH LINEAR PRIMERS - The present disclosure relates to the amplification of target nucleic acid sequences for various sequencing and/or identification techniques. The use of these primers, as described herein, allows for the reduction in the amplification of nonspecific hybridization events (such as primer dimerization) while allowing for the amplification of the target nucleic acid sequences.11-26-2009
20090280539DNA POLYMERASES AND RELATED METHODS - Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. The mutant polymerases overcome the inhibitory effects by an intercalating dye. Therefore, the mutant polymerases are useful in a variety of disclosed methods in combination with an intercalating dye. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.11-12-2009
20100159528POLYMERASE STABILIZATION - The present invention relates to methods and compositions for providing purified thermostable enzymes, particularly thermostable DNA polymerases, that are free of exogenous detergents. The present invention also provides methods for providing such purified thermostable DNA polymerases to assays in an active form by adding one or more detergents. The present invention further provides compositions and kits comprising purified thermostable DNA polymerases for use in a variety of applications, including amplification and sequencing of nucleic acids.06-24-2010
20080305528ISOLATION OF NUCLEIC ACIDS - A method for extracting nucleic acids from a biological material such as blood comprises contacting the mixture with a material at a pH such that the material is positively charged and will bind negatively charged nucleic acids and then eluting the nucleic acids at a pH when the said materials possess a neutral or negative charge to release the nucleic acids. The nucleic acids can be removed under mildly alkaline conditions to the maintain integrity of the nucleic acids and to allow retrieval of the nucleic acids in reagents that are immediately compatible with either storage or analytical testing.12-11-2008
20100221785Isothermal Strand Displacement Amplification Using Primers Containing a Non-Regular Base - The invention is directed to a method for isothermal DNA amplification comprising providing to the DNA to be amplified an amplification mix comprising a first primer at least partially complementary to a region of DNA and containing a non-regular base, a second primer at least partially complementary to a region of DNA and containing a non-regular base, a DNA polymerase, an enzyme capable of strand displacement, an enzyme that recognises a non-regular base in double-stranded DNA and causes a nick or excises a base in one DNA strand at or near the non-regular base; and amplifying the DNA substantially without thermal cycling.09-02-2010
20110136179MICROFLUIDIC BIOCHIP - The present invention provides a microfluidic biochip, which comprises: a fluid transportation unit having a fluid transportation reservoir and a fluid transportation air chamber; a first fluid storage reservoir; a second fluid storage reservoir; a first valve unit having a first valve and a first valve control air chamber; and a second valve unit having a second valve and a second valve control air chamber; wherein the first valve unit is located between the first fluid storage reservoir and the fluid transportation unit, the second valve unit is located between the second fluid storage reservoir and the fluid transportation unit, and the top portion of the fluid transportation reservoir and the valves are made of a flexible material. The structure of the present microfluidic biochip allows fluids to be transported and/or mixed therein.06-09-2011
20090035823Ligation-based synthesis of oligonucleotides with block structure - The present invention relates to a method of producing single-stranded nucleic acid molecules from oligo- or polynucleotides wherein each of said oligo- or polynucleotides has a predefined 5′ or 3′ terminus, comprising the steps of (a) annealing an adaptor oligonucleotide simultaneously or step by step to (aa) a first oligo- or polynucleotide; and (ab) a second oligo- or polynucleotide wherein the 5′-terminus of said adaptor oligonucleotide is complementary in sequence to the 5′ terminus of said first oligo- or polynucleotide and the 3′terminus of said adaptor molecule is complementary in sequence to the 3′ terminus of said second oligo- or polynucleotide; and optionally (a′) simultaneously with or subsequently to step (a) annealing at least one further adaptor oligonucleotide to free termini of said first or second oligonucleotides and to free termini of further oligo- or polynucleotides; (b) optionally filling in gaps between the neighbouring ends of said oligo- or polynucleotides; (c) ligating said oligo- or polynucleotides; and (d) removing said at least one adaptor oligonucleotide. In a preferred embodiment of the method of the invention, said single-stranded nucleic acid molecules represent a collection of nucleic acid molecules wherein either said first or said second oligo- or polynucleotide is invariable in sequence between all members of said collection of nucleic acid molecules.02-05-2009
20110300583IN VITRO RECOMBINATION METHODS USING 5' EXONUCLEASE - Provided herein is a method for homologous end cloning using a 5′-exonuclease. One aspect provides a method of generating a recombinant DNA molecule comprising 5′ exonuclease digestion of DNA molecules, exonuclease inactivation, annealing of 3′ overhangs to form a annealed complex. Also provided is a method a method of generating a recombinant DNA molecule comprising 5′ exonuclease digestion of DNA molecules, exonuclease inactivation, annealing of 3′ overhangs to form a annealed complex, and extension of 3′ ends to fill gaps corresponding to exonuclease removal to form a substantially non-gapped annealed complex. Also provided is a one reaction method in which no further additions need be made to the reaction mixture and the reaction mixture is only manipulated thereafter as to incubation time and incubation temperature. Complexes formed according to methods described herein can be transformed into a host cell without further in vitro processing.12-08-2011
20120107878MULTIPLEX ASSEMBLY OF HIGH FEDELITY DNA - The present invention relates to novel sequence normalization protocols and methods which utilize this protocol to provide a robust multiplexed assembly of high fidelity polynucleotides/genes05-03-2012
20110287489RECOMBINATIONAL CLONING USING ENGINEERED RECOMBINATION SITES - Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).11-24-2011
20110294167NUCLEIC ACID AMPLIFICATION - The present invention provides improved systems and methods for amplifying nucleic acids. Among other things the present invention provides a system for amplifying nucleic acids through use of a primase and a polymerase with strand-displacement ability without, for example, exogenously-added primers. The present invention is particularly useful for whole genome amplification.12-01-2011
20090298129SYSTEMS AND METHODS FOR PROCESSING SAMPLES IN A CLOSED CONTAINER, AND RELATED DEVICES - A system and method for automated processing of nucleic acids and other samples includes a disposable container comprising a tray and a flexible barrier. The barrier is configured to seal with a top edge of the tray, providing a closed, aseptic work area within the sealed tray. A pipette head and/or other sample manipulation device can be attached to the inside of the barrier, and the barrier can include an interface for a robotic arm or other device. When the barrier is sealed over the tray, the barrier separates the contents of the tray from the robot or other manipulation device. The barrier can be flexible, and allow the robotic arm to move the pipette head throughout the work area of the tray. All samples, reagents, pipette tips and other tools or devices for processing nucleic acid samples may remain within the closed compartment provided by the container during processing.12-03-2009
20080318281Thermostabillization of Dna Polymerase by Protein Folding Pathway from a Hyperthermophile Archaeon, Pyrococcus Furiosus - The present invention relates to maintaining the activity and stability of enzymes and biologically active proteins at increased temperatures by contacting same with a combination of isolated passive and active chaperones from a hyperthermopilic Archaeon, wherein the chaperones may include heat shock proteins, prefoldin and/or chaperonin proteins.12-25-2008
20100267092COMPONENTS - A method and device structure are provided which enable an archive sample to be collected and detached relative to a device within which a series of processes, such as PCR are being provided. A chamber structure and method of use are provided in which a controlled and precise volume is obtained by control of the relative resistance to flow through various channels.10-21-2010
20090098613Method for synthesizing DNA - A DNA synthesis reaction-enhancer comprising at least one kind selected from the group consisting of acidic substances and cationic complexes; a DNA synthesis method in which during a DNA synthesis reaction a reaction is carried out in the presence of the above enhancer by using DNA polymerase; a DNA synthesis reaction composition comprising the above enhancer; a DNA synthesis reaction composition comprising two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity; a DNA synthesis method in which during a DNA synthesis reaction two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity are used; a kit for use in in vitro DNA synthesis, comprising two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity; and a kit for use in in vitro DNA synthesis, wherein the kit comprises the DNA synthesis reaction-enhancer and DNA polymerase. According to the present invention, DNA synthesis can be carried out at an efficiency more excellent as compared to conventional DNA synthesis reaction.04-16-2009
20090098612METHOD OF AMPLIFYING A TARGET NUCLEIC ACID BY ROLLING CIRCLE AMPLIFICATION - Provided is kit for and a method of amplifying a nucleic acid using rolling cyclic amplification (RCA), including amplifying a nucleic acid together with formation of a single-strand circular DNA template using RCA by reacting a reaction solution including: (a) two hairpin oligos, (b) a target nucleic acid, (c) a DNA ligase,(d) an endonuclease, (e) a DNA polymerase, and (f) a primer.04-16-2009
20080311627Compositions and Methods for Preventing Carry-Over Contamination in Nucleic Acid Amplification Reactions - Particular aspects provide methods for the specific amplification of template DNA in the presence of potentially contaminating PCR products from previous amplification experiments, but wherein the contaminating prior reaction amplificates (carry-over contaminants) are rendered non-amplifiable, based on use of particular contaminant degradation enzymes, and use of at least one primer that is fully complementary to any contaminating prior reaction amplificate nucleic acid, but contains a mismatch with the correspond sequence of the sample template nucleic acid. Additional aspects provide a method for the specific amplification of single-stranded sample template DNA in the presence of potentially double-stranded carry-over products. Further aspects provide methods comprising use of a template-dependent thermostable DNA polymerase enzyme suitable for incorporating ribonucleotides as well as deoxy-nucleotides to provide a chimeric amplificate. After digestion with an RNase, any contaminating prior chimeric amplificate, the RNase is inactivated. These aspects are surprisingly effective alternatives to the carry over protection system known as UNG system, and other art-recognized methods.12-18-2008
20080311626Dna Polymerases Having Strand Displacement Activity - The invention provides novel strand displacement DNA polymerases which can be used in a rapid and efficient strand displacement amplification reactions. The polymerases are significantly more thermostable than prior art polymerase and retain high activity at elevated temperatures. Also disclosed are genes encoding the polymerases and vectors comprising the genes. Representative polymerases of the invention are obtainable from bacterial strains of the species 12-18-2008
20080311629Development of Pcr Primers and Primer Mixtures For Amplification of Cnp60 Target Sequences - We have developed the primer pair H1511 and H1261 as a replacement for primer pair H279/H280 for specific amplification of cpn60 universal target sequences in genomic DNA or in complex DNA mixtures, including those with high G+C content.12-18-2008
20100035303Method of Amplifying Target Nucleic Acid Sequence By Multiple Displacement Amplification Including Thermal Cycling - A method of amplifying a target nucleic acid sequence includes multiple displacement amplification and thermal cycling. According to the method, the target nucleic acid sequence may be effectively amplified.02-11-2010
20100112644Methods for detection and quantitation of small RNAs - Improved methods that increase the specificity and sensitivity of detection of small RNAs, including miRNAs, using oligonucleotide primers and nucleic acid amplification, are provided. Reaction conditions that result in preferential decrease in cDNA synthesis of RNAs other than the small RNA molecules targeted for detection during miRNA tailing and reverse transcription reactions are described. Using these reaction conditions greater sensitivity and specificity of amplification of small RNAs including miRNAs is achieved.05-06-2010
20100112643METHOD FOR DIRECT CAPTURE OF RIBONUCLEIC ACID - A method of: providing a solid surface having a dendrimer molecule bound thereto and a single-stranded probe nucleic acid immobilized to the dendrimer; contacting the solid surface with a sample suspected or known to contain a target ribonucleic acid; denaturing the target ribonucleic acid; and incubating the sample to allow hybridization of the denatured ribonucleic acid to the probe nucleic acids. The target ribonucleic acid is complementary to the probe nucleic acid.05-06-2010
20090148911DNA POLYMERASES WITH ENHANCED LENGTH OF PRIMER EXTENSION - A formulation and kit of thermostable or other DNA polymerases comprising at least one thermostable or other DNA polymerase which lacks 3′-exonuclease activity, and at least one thermostable DNA polymerase exhibiting 3′-exonuclease activity. Also provided is an improved method for enzymatic extension of DNA strands, especially while, but not limited to, amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is made and used to catalyze primer extension.06-11-2009
20100267091Method for a continuous rapid thermal cycle system - Disclosed herein is an efficient, high speed production scale synthesis method for high molecular weight organic substances, such as DNA. The invention includes a method of conducting a polymerase chain reaction which comprises transporting a liquid through polymeric tubing disposed through a first reaction cycle region and at least a second reaction cycle region, each of which regions comprises at least a first and a second temperature zone, the temperature in each zone of said at least second region being substantially identical to the corresponding first and second zones in said first region, wherein said liquid is an aqueous solution comprising polymerase chain reaction reactants and a surface absorbing polymer.10-21-2010
20100279358METHODS FOR NUCLEIC ACID MANIPULATION - A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.11-04-2010
20100124766Apparatus and Method for Segmented Thermal Cycler - The present invention relates to a thermal cycler for the carrying out of chemical or biological reactions, such as PCR or other nucleic acid amplification reactions, that is segmented with a plurality of reaction vessel receiving elements. The reaction vessel receiving elements are thermally isolated from each other and provide an airtight seal to prevent liquids or moisture from penetrating below the reaction vessel receiving elements. The reaction vessel receiving elements have several recesses arranged in a pattern to receive the reaction vessels of a single standard microtiter plate and the segmented thermal cycler has a system for independently heating and cooling each of the reaction vessel receiving elements.05-20-2010
20120142059SEQUENCE AMPLIFICATION WITH TARGET PRIMERS - The present disclosure relates to the amplification of target nucleic acid sequences for various sequencing and/or identification techniques. This can be accomplished via the use of target primers and isothermal multiple strand displacement (MDA) processes. The use of these target primers and MDA, as described herein, allows for the reduction in the amplification of undesired hybridization events (such as primer dimerization and the “jackpot mutation” effect of PCR) while allowing for the amplification of the target nucleic acid sequences.06-07-2012
20100081175ENZYME-CONTAINING GELS AND NUCLEIC ACID AMPLIFYING KITS - The present invention is aimed at providing an enzyme-containing gel which is employed for deactivating, from a sample containing proteins and nucleic acids, easily and simply the proteins in the sample and then amplifying a nucleic acid in the sample, a kit for the amplification of nucleic acid which comprises the enzyme-containing gels, and a method for the amplification of nucleic acid which employs the enzyme-containing gels. The present invention provides an enzyme-containing gel which is characterized in that the gel contains a heat-resistant enzyme; a kit for the amplification of nucleic acid, comprising the above-described enzyme-containing gels and a proteolytic enzyme; and a method for the amplification of nucleic acid, employing the above-described enzyme-containing gels.04-01-2010
20100081176ENZYME-CONTAINING CAPSULES AND NUCLEIC ACID AMPLIFICATION KITS - An object of the present invention is to provide an enzyme-containing capsule which is employed for deactivating, from a sample, the proteins in the sample and then amplifying a nucleic acid in the sample, a kit for the amplification of nucleic acid which comprises the enzyme-containing capsules, and a method for the amplification of nucleic acid which employs the enzyme-containing capsules. The present invention provides an enzyme-containing capsule which is characterized in that the capsule has a melting point of 60 to 95° C., comprises a non-proteinous material as the envelope component, and contains a heat-resistant enzyme in the inside of the capsule; a kit for the amplification of nucleic acid, comprising the above-described enzyme-containing capsules and a proteolytic enzyme; and a method for the amplification of nucleic acid, employing the above-described enzyme-containing capsules.04-01-2010
20090263869Methods and Compositions for PCR - A modified thermostable Pol B DNA polymerase, produced by a reaction, under essentially aqueous conditions, of a thermostable Pol B DNA polymerase and a modifier reagent of Formula I wherein the reaction results in a thermally reversible inactivation of the thermostable Pol B DNA polymerase activity and the 3′-5′ exonuclease activity, which polymerase is suitable for hot-start PCR. Also disclosed are the method for the modification, a polynucleic acid amplification method and PCR reaction mixture and kit comprising the modified thermostable Pol B DNA polymerase.10-22-2009
20110201056Repair of Nucleic Acids for Improved Amplification - Methods and compositions are provided for repairing a polynucleotide so that it can be synthesized efficiently with improved fidelity and yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NAD+ or ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI.08-18-2011
20120295312ROTATIONAL PCR EQUIPMENT AND PCR METHOD USING THE SAME - A rotational PCR apparatus, a PCR chip for the same and a rotational PCR method using the same.11-22-2012
20090170167SINGLE ENZYME SYSTEM FOR FAST, ULTRA LONG PCR - The present invention provides methods, formulation and kits for the synthesis of long nucleic acid fragments. An improved PCR method is provided for amplifying long DNA fragments. In particular, a single thermostable DNA polymerase is used for the rapid amplification of over 10 kb long DNA fragments. Also provided is a method for extending long complementary DNA strands using this single enzyme formulation.07-02-2009
20100124765SEQUENCE AMPLIFICATION WITH TARGET PRIMERS - The present disclosure relates to the amplification of target nucleic acid sequences for various sequencing and/or identification techniques. This can be accomplished via the use of target primers. The use of these target primers, as described herein, allows for the reduction in the amplification of undesired hybridization events (such as primer dimerization) while allowing for the amplification of the target nucleic acid sequences.05-20-2010
200901428102'-Terminator Nucleotide-Related Methods and Systems - The present invention provides methods of extending primer nucleic acids and sequencing target nucleic acids. The methods include the use of 2′-terminator nucleotides to effect chain termination. In addition to related reaction mixtures and kits, the invention also provides computers and computer readable media.06-04-2009
20090280537METHODS AND COMPOSITIONS FOR DETECTING DIHYDROPYRIMIDINE DEHYDROGENASE SPLICING MUTATIONS - The present invention provides methods, compositions and kits for the detection of genetic polymorphisms or mutations of the dihydropyrimidine dehydrogenase deficiency (DPDD). The polymorphisms or mutations generally occur in the dihydropyrimidine dehydrogenase (DPD) gene in chromosome 1. Also provided are mutant forms of DPD.11-12-2009
20090280540DIRECTED ENRICHMENT OF GENOMIC DNA FOR HIGH-THROUGHPUT SEQUENCING - The present invention provides microarrays of oligonucleotide primer pairs, and in particular, microarrays of primers that comprise at least one cleavable linkage. Also provided are methods to capture oligonucleotide primer pairs from one or more microarrays, and methods to use the captured oligonucleotide primer pairs, such as for amplification of a target polynucleotide sequence. In addition, methods of using a microarray to isolate, purify and/or amplify a target polynucleotide are provided.11-12-2009
20090286285Modified Nucleic Acid Polymers and Methods for their Production - The invention is directed to nucleic acid sequences having decreased thermodynamic stability to complementary sequence as well as a method for producing these sequences.11-19-2009
20100136631IMPROVED PRIMERS AND PROBES FOR THE AMPLIFICATION AND DETECTION OF HIV GAG,REV AND NEF - The invention relates to improved methods and compositions for the nucleic acid amplification of one or multiple variants (strains) of Human Immunodeficiency Virus (HIV) present in a sample, and preferably in a sample from a pathogen infected individual. In particular, novel primers, methods and kits for the amplification of one or more species of HIV Rev, Gag and Nef nucleic acids are provided. The amplified HIV nucleic acid can be used to identify and/or quantitate HIV variants present in a sample. Nucleic acids produced by the methods of the invention or the proteins encoded thereby can also be used directly as vaccines or to transfect/load antigen presenting cells. The loaded antigen presenting cells can be used as a vaccine for the treatment or prevention of HIV infection.06-03-2010
20090298131Non-Emulsion Methods And Masked Biomolecules - The present invention relates to methods and devices for amplifying nucleic acid, and, in particular, amplifying so as to generate products on a surface without the use of emulsions. In a preferred embodiment, a plurality of groups of amplified product are generated on the surface, each group positioned in different (typically predetermined) locations on said surface so as to create an array.12-03-2009
20090298130LADDER ASSEMBLY AND SYSTEM FOR GENERATING DIVERSITY - The present invention provides novel methods of generating a nucleic acid molecule. In certain embodiments, a double stranded nucleic acid chunk is generated from a ladder complex comprising partially complementary oligonucleotides, which chunk is combined with a nucleic acid acceptor molecule. In certain embodiments, the assembled chunk/nucleic acid acceptor molecule complex may be propagated in vivo or in vitro. The present invention also provides improved systems for generating a plurality of nucleic acid molecules that differ at one or more nucleotide positions. In certain embodiments, the plurality of nucleic acid molecules encodes a polypeptide or portion of a polypeptide.12-03-2009
20120295311Methods and Materials for Nucleic Acid Manipulation - The present invention is concerned with the field of nucleic acid manipulation and particularly DNA manipulation, and uses thereof. Specifically, the invention pertains to methods involving the joining of nucleic acids and uses of such joined nucleic acids, for example for creating transformed microorganisms. Also, the invention pertains to materials useful in such methods.11-22-2012
20090170166ANTIBODY INHIBITING URACIL-DNA-GLYCOSYLASE AND USES THEREOF FOR DECONTAMINATING NUCLEIC ACID AMPLIFICATION REACTIONS - The invention concerns antibodies directed against the uracil-DNA-glycosylase inhibitor, and uses thereof for decontaminating nucleic acid amplification reactions.07-02-2009
20090035824NUCLEIC ACID-TEMPLATED CHEMISTRY IN ORGANIC SOLVENTS - The present invention provides methods and compositions for performing nucleic acid mediated chemistry in a variety of organic solvents. A variety of nucleic acid mediated chemical reactions may be efficiently carried out in organic solvents.02-05-2009
20080274513Method and Device for Conducting Biochemical or Chemical Reactions at Multiple Temperatures - Methods and devices for conducting chemical or biochemical reactions that require multiple reaction temperatures are described. The methods involve moving one or more reaction droplets or reaction volumes through various reaction zones having different temperatures on a microfluidics apparatus. The devices comprise a microfluidics apparatus comprising appropriate actuators capable of moving reaction droplets or reaction volumes through the various reaction zones.11-06-2008
20080274511DEVICE FOR CARRYING OUT CHEMICAL OR BIOLOGICAL REACTIONS - The invention relates to a device for carrying out of chemical or biological reactions with a reaction vessel receiving element for receiving a microtiter plate with several reaction vessels, wherein the reaction vessel receiving element has several recesses arranged in a regular pattern to receive the respective reaction vessels, a heating device for heating the reaction vessel receiving element, and a cooling device for cooling the reaction vessel. The invention is characterized by the fact that the reaction vessel receiving element is divided into several segments. The individual segments are thermally decoupled from one another, and each segment is assigned a heating device which may be actuated independently of the others. By means of the segmentation of the reaction vessel receiving element, it is possible for zones to be set and held at different temperatures. Because the reaction vessel receiving element is suitable for receiving standard microtiter plates, the device according to the invention may be integrated in existing process sequences.11-06-2008
20080286835Isothermal Amplification of Nucleic Acids - A process of amplifying a nucleic acid template dependent on partial destruction of primer molecules which have extended onto the template molecule followed by strand invasion of the partially destroyed primer template by a replacement primer. The destruction of the primer molecule may be performed by either endonuclease or exonuclease digestion. A signal generation from the amplified products may be obtained by the use of adaptors capable of binding probe molecules as well as the amplified product.11-20-2008
20080293107METHODS FOR USING RIBOPRIMERS FOR STRAND DISPLACEMENT REPLICATION OF TARGET SEQUENCES - Methods, compositions and kits for amplifying a target sequence by strand displacement replication using strand-displacing primers. The method uses primers that have only ribonucleotides or purine ribonucleotides and at least one 2′-substituted pyrimidine-2′-deoxyribonucleotide.11-27-2008
20080280331Microfluidic Analysis System - A thermal cycling device (11-13-2008
20100279357METHODS FOR NUCLEIC ACID MANIPULATION - A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.11-04-2010
20080261277pRNA chimera - A circularly permuted chimeric pRNA molecule carrying a stabilized biologically active RNA, such as a ribozyme.10-23-2008
20090017505Process and device for mixing microdroplets - In a process for mixing microdroplets (01-15-2009
20110207180TEMPERATURE CONTROL DEVICE WITH A FLEXIBLE TEMPERATURE CONTROL SURFACE - A device for controlling temperature in a reaction chamber is disclosed. The device comprises: a bladder assembly comprising a housing dimensioned to hold a reaction chamber disposed within an interior volume of the housing; and a first temperature-control bladder disposed within the housing, the first temperature-control bladder is configured to receive a temperature-control fluid and comprises a flexible, heat conductive surface that comes in contact with at least a portion of an exterior surface of the reaction chamber after receiving the temperature-control fluid. Also disclosed are a bladder thermal cycler, a temperature-control bladder assembly and methods for producing a thermal cycle in a reaction chamber.08-25-2011
20080213842KIT FOR SYNTHESIZING POLYNUCLEOTIDES - The present invention realized isothermal and rapid polynucleotide synthesis by using as templates polynucleotides having a structure capable of forming loops, and combining a plurality of primers capable of providing a starting point for complementary strand synthesis to such loops. If the LAMP method is applied, all reactions can be carried out isothermally and rapidly since the template polynucleotides themselves can also be synthesized by an isothermal reaction.09-04-2008
20100143979POLYPEPTIDES HAVING DNA POLYMERASE ACTIVITY - A polypeptide having a high fidelity DNA polymerase activity and thus being useful as a genetic engineering reagent; a gene encoding this polypeptide; a method of producing the polypeptide; and a method of amplifying a nucleic acid by using the polypeptide.06-10-2010
20080268506Reaction Vessel - A reaction vessel for use in thermal cycling reactions is disclosed. The vessel has a width greater than its depth, to give the vessel a flattened profile. The side walls of the vessel extending across the width of the vessel are generally flat, and have a lesser thickness than the side walls extending across the depth of the vessel, This conformation allows rapid heat transfer into the vessel during thermal cycling, while the flattened profile allows optical detection techniques to be used on the contents of the vessel. The tubes may be provided singly or in an array or multiwell formal. Also described is a carousel for holding the tubes for use in a thermal cycler.10-30-2008
20080268505Method for creating polynucleotide and polypeptide sequences - The invention provides methods for evolving a polynucleotide toward acquisition of a desired property. Such methods entail incubating a population of parental polynucleotide variants under conditions to generate annealed polynucleotides comprising heteroduplexes. The heteroduplexes are then exposed to a cellular DNA repair system to convert the heteroduplexes to parental polynucleotide variants or recombined polynucleotide variants. The resulting polynucleotides are then screened or selected for the desired property.10-30-2008
20100136632FULLY AUTOMATED PORTABLE DNA DETECTION SYSTEM - Provided herein is a portable thermocycler, comprising: (i) a case; (ii) a rotary plate in the case; (iii) a plurality of heating blocks arranged in a geometric pattern disposed on the rotary plate; and (iv) at least one vessel adapted to move and contact at least two of the plurality of heating blocks; wherein each of the heating blocks comprises a heating plate maintained at a set temperature over a thermally insulating material; wherein the geometric pattern comprises a number of center heating blocks arranged in a shape defining a polygon and a number of outside heating blocks disposed around the periphery of the rotary plate; and wherein the rotary plate includes a plurality of rotating wheels adapted to rotate at least one of the vessels into contact with each of the heating blocks.06-03-2010
20100273218COMPOSITIONS AND METHOD FOR STORAGE OF NUCLEIC ACID FROM BODILY FLUIDS - The present invention provides an aqueous composition comprising SDS, Cyclohexanediamine tetraacetate, Tris-HCl and proteinase K for the extraction of nucleic acid from a sample of bodily fluid, such a saliva, wherein the extracted nucleic acid is stable for at least fourteen days at room temperature The composition permits direct use of the extracted and stored DNA in an amplification reaction without further processing.10-28-2010
20090047714OLIGONUCLEOTIDES AS TEMPERATURE-SENSITIVE INHIBITORS FOR DNA POLYMERASES - Aspects of the invention relate to the use of novel oligonucleotides as temperature-sensitive inhibitors for thermostable DNA polymerases. Some inhibitors exhibit temperature-dependent and, in some cases, reversible inhibitory property by changing the conformation of at least a portion of the oligonucleotides from double-stranded to single stranded or in some cases vice versa in a temperature-dependent manner. Aspects also relate to the use of an the inhibitors in a hot-start PCR compositions, wherein the inhibitor may act to suppress the activity of the thermostable DNA polymerase below a desired activation temperature, Tact, and wherein the inhibitor is thermally inactivated above Tact, thus liberating the polymerase activity and initiating the DNA amplification process. Aspects further relate to a procedure for formulating the composition of a hot-start PCR reaction mixture. The hot-start PCR methods disclosed herein are generally faster, more flexible and lower in cost than existing methods.02-19-2009
20110269192LOOP-SHAPED PRIMER USED IN NUCLEIC ACID AMPLIFICATION AND THE USE THEREOF - Loop-shaped primer used in nucleic acid amplification is an oligonucleotide with 3-20 bases in both 3′ and 5′ ends which can be combined together to form a double-strand under appropriate conditions, resulting in the primer forming a stem-loop structure. The double-stranded structure is opened and the stem-loop structure dissolves when the primer recognizes and hybridizes with the target sequence. If the target sequence is not present the primer can form a stem-loop structure automatically by self-annealing. The primer can comprise a universal tag sequence or not. Together with universal tag sequence primer the primer comprising the universal tag sequence can be used for a second round of amplification. The primer has high specificity and does not form a primer dimmer. The primer is easy to design and is suitable for measuring gene expression and detecting features of nucleic acids such as SNPs and rare mutations.11-03-2011
20090286286Methods for controlling amplification - Methods of amplifying nucleic acid on a solid support are described. Beads and template, each in known concentrations, are employed so a range of template to bead ratios can be exploited. Where the beads contain primers, the template can be amplified. After amplification, non-covalently bound template is removed, so as to leave beads with extended primers (or beads with primers that were not extended).11-19-2009
20090137007Method for sequencing nucleic acid molecules - The present invention is directed to a method of sequencing a target nucleic acid molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the template nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.05-28-2009
20090186387RECOMBINATIONAL CLONING USING ENGINEERED RECOMBINATION SITES - Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).07-23-2009
20080318280COVER FOR AN ARRAY OF REACTION VESSELS FOR ONE-STEP OPERATION MODUS - The present invention relates to means for covering one or more sample(s) that are suitable to avoid or minimize evaporation and/or condensation of any vaporizable substance that may be present in the sample(s) or reaction mixture(s), in particular evaporation of substance at the fringes of a vessel or an array of vessels or condensation of said substance on the lid of a reaction vessel or a plate/block containing the sample(s) and/or the means for covering. This is achieved by providing a device comprising, among others, a means for positioning a covering means and a means for guiding at least one lid of the covering means.12-25-2008
20090325237Method of Analyzing a BRCA2 Gene in a Human Subject - Five novel DNA and protein sequences have been determined for the BRCA2 gene, as have been ten polymorphic sites and their rates of occurrence in the normal alleles of BRCA2. The sequences BRCA212-31-2009
20090325239METHODS FOR NUCLEIC ACID MAPPING AND IDENTIFICATION OF FINE-STRUCTURAL-VARIATIONS IN NUCLEIC ACIDS - A method of juxtaposing sequence tags (GVTs) that are unique positional markers along the length of a population of target nucleic acid molecules is provided, the method comprising: fragmenting the target nucleic acid molecule to form target DNA insert; ligating the target DNA insert to a DNA vector or backbone to create a circular molecule; digesting the target DNA insert endonuclease to cleave the target DNA insert at a distance from each end of the target DNA insert yielding two GVTs comprising terminal sequences of the target DNA insert attached to an undigested linear backbone; recircularizing the linear backbone with the attached GVTs to obtain a circular DNA containing a GVT-pair having two juxtaposed GVTs; and recovering the GVT-pair DNA by nucleic acid amplification or digestion with endonuclease having sites flanking the GVT-pair. Cosmid vectors are provided for creating GVT-pairs of ˜45- to 50-kb separation sequencable by next-generation DNA sequencers.12-31-2009
20090325236Optical sorting method - The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising the steps of: (a) compartmentalising genetic elements into microcapsules; (b) expressing the genetic elements to produce their respective gene products within the microcapsules; (c) sorting the genetic elements which produce the gene product having the desired activity using a change in the optical properties of the genetic elements. The invention enables the in vitro evolution of nucleic acids and proteins by repeated mutagenesis and iterative applications of the method of the invention.12-31-2009
20090325234Apparatus and method for a continuous rapid thermal cycle system - A thermal cycle system and method suitable for mass production of DNA comprising a temperature control body having at least two sectors. Each sector has at least one heater, cooler, or other means for changing temperature. A path traverses the sectors in a cyclical fashion. In use, a piece of tubing or other means for conveying is placed along the path and a reaction mixture is pumped or otherwise moved along the path such that the reaction mixture is repetitively heated or cooled to varying temperatures as the reaction mixture cyclically traverses the sectors. The reaction mixture thereby reacts to form a product. In particular, polymerase chain reaction reactants may continuously be pumped through the tubing to amplify DNA. The temperature control body is preferably a single aluminum cylinder with a grooved channel circling around its exterior surface, and preferably has wedge-shaped or pie-shaped sectors separated by a thermal barrier.12-31-2009
20090203083SUBSTRATE FOR NUCLEIC ACID AMPLIFICATION - The present invention relates to a method, a substrate, a kit, and a system for nucleic acid amplification comprising a porous substrate with pores enabling the diffusion of biomolecules. More particular, the present invention relates to a method, a substrate, a kit and a system, wherein the nucleic acid amplification takes place within the pores of a porous substrate.08-13-2009
20110143399DNA JOINING METHOD - The present invention provides a method to directionally clone any linear template DNA molecule into any linearized vector. The vector ends may be generated from any restriction enzyme cleavage. The method does not require a ligation step nor the use of carefully controlled conditions as is required with methods involving specific exonucleases alone. It has been determined that specific DNA polymerases are able to efficiently join one or more linear DNA molecules sharing ends with appropriate complementation.06-16-2011
20100003724CHEMICALLY MODIFIED NUCLEOSIDE 5'-TRIPHOSPHATES FOR THERMALLY INITIATED AMPLIFICATION OF NUCLEIC ACID - Provided herein are methods and compositions for nucleic acid replication. These methods involve the use of 3′-substituted nucleoside 5′-triphosphates or 3′-substituted terminated primers in nucleic acid replication reactions. In certain aspects, the methods are accomplished by use of 3′-substituted NTPs and/or 3′-substituted terminated primers which provide utility in nucleic acid replication. In preferred embodiments, the NTPs and/or primers are substituted at the 3′-position with particular heat labile chemical groups such as ethers, esters or carbonate esters.01-07-2010
20090087884Microfluidic nucleic acid amplification and separation - Microfluidic devices designed for assaying biochemical molecules are disclosed. The microfluidic devices are capable of assaying nucleic acids for identification of nucleic acid species. The microfluidic devices are adapted to carry out an amplification of the nucleic acid and subsequent separation of amplified nucleic acid species. Also disclosed is a method for amplifying and separating a nucleic acid sample on a microfluidic device.04-02-2009
20090191595Method for amplification of long nucleic acid - An object of the present invention is to provide a method for amplification of long nucleic acid, wherein the method allows nucleic acid fragments containing the same nucleotide sequence information to efficiently amplify at the same base length. The present invention relates to a method for amplification of long nucleic acid sequence, wherein the method uses primers being modified at the 5′ end with a phosphate group and performs a cooperative reaction using DNA polymerase and DNA ligase.07-30-2009
20090191596ALKALINE SHOCK-BASED PREPARATION OF NUCLEIC ACIDS - Method of preparing a biological sample appropriate for use in a subsequent in vitro nucleic acid amplification reaction. The method involves a rapid, transient exposure to alkaline conditions which can be achieved by mixing an alkaline solution with a pH-buffered solution that includes a detergent and the biological sample to be tested for the presence of particular nucleic acid species using in vitro amplification. The invented method advantageously can improve detection of some target nucleic acids without substantially compromising detectability of others. The method is particularly useful for simultaneously preparing RNA and DNA templates that can be used in multiplex amplification reactions.07-30-2009
20090191594Microchemistry reaction method and device - A chemical reaction is conducted in a fluid of a droplet inside a reaction receptacle or on a surface of a reaction substrate. Fluctuations of a magnetic field are applied to the droplet including an aqueous solution having magnetic body particles with a hydrophilic surface, and a physical force is transmitted to the surrounding aqueous solution through the magnetic body particles. The droplet is thus moved by the physical force to conduct an operation necessary for a chemical reaction.07-30-2009
20090053773Reaction Container and Dna Amplification Reaction Method - A linear recess is provided in a reverse side surface of a synthetic resin base plate 02-26-2009
20090053772Thermal Cycler for PCR Including Temperature Control Bladder - Methods and devices for performing chemical reactions under controlled temperatures are described. In one embodiment, the devices provided by the invention comprise a housing dimensioned to hold a reaction chamber disposed within an interior volume of the housing. The reaction chamber has thermally conductive interior and exterior surfaces defining an internal volume therein at a first temperature. The device also includes at least one thermally conductive temperature-control bladder disposed therein, which bladder is configured to receive a temperature-control substance at a second temperature into said bladder and expel said temperature-control substance from said bladder. The bladder is further configured such that upon receiving the temperature-control substance, the bladder expands to abut substantially at least a portion of said exterior surfaces of said reaction chamber to enable thermal exchange between said temperature-control substance the said internal volume of reaction chamber.02-26-2009
20090011470Nucleic acid sample preparation by exclusion of DNA - Devices and methods are provided for separation of nucleic acids from waste materials in a biological sample, and then reacting the separated nucleic acids. In some embodiments, the methods comprise mixing a cell lysate having nucleic acids and waste materials with a waste-binding matrix to capture the cellular waste materials from the lysate. The waste-binding matrix can comprise hydrophilic and/or hydrophobic size-exclusion, ion-exchange particles. In some embodiments, the device comprises a substrate comprising a fluid processing pathway in which nucleic acid sample preparation occurs prior to a downstream genotyping reaction.01-08-2009
20080318282Nucleic acid amplification method - Provided is a simple and highly sensitive nucleic acid amplification method including hybridizing two types of oligonucleotide probes with a target gene and ligating the oligonucleotide probes with DNA ligase and amplifying the resultant single-stranded oligonucleotide in accordance with LAMP.12-25-2008
20090203085Isothermal Nucleic Acid Amplification Methods and Compositions - Methods and compositions are provided related to the amplification of target polynucleotide sequences as well as total RNA and total DNA amplification. In some embodiments, the methods and compositions also allow for the immobilization and capture of target polynucleotides with defined 3′ and or 5′ sequences to solid surfaces. The polynucleotides attached to the solid surfaces can be amplified or eluted for downstream processing. In some cases, nucleotides attached to solid surfaces can be used for high throughput sequencing of nucleotide sequences related to target DNA or target RNA.08-13-2009
20090203084METHODS AND APPARATUSES FOR CONVECTIVE POLYMERASE CHAIN REACTION (PCR) - The present invention provides a method and apparatus for amplifying a nucleic acid sequence by polymerase chain reaction (PCR). The method comprises placing a PCR sample in a container which is heated by only a single heat source that provides a high temperature for denaturation in the bottom of the PCR sample, while annealing and extension automatically occur in different regions of the PCR sample due to the convection induced by a temperature gradient descending from the bottom of the PCR sample to the surface of the PCR sample.08-13-2009
20090203082Thermocycling of a Block Comprising Multiple Sample - The present invention relates to the field of high throughput analysis of samples. In particular, the present invention is directed to a device, a System and a method for simultaneous tempering of multiple samples. More particular, the invention relates to the simultaneous thermocycling of multiple samples to perform PCR in a microtiter plate format.08-13-2009
20090004701Multiplex oligonucleotide addition and target amplification - Methods for appending oligonucleotides directly to nucleic acid templates, particularly to defined sites internal to single-stranded templates, are described. Appending first and second common priming sites to each of a plurality of templates of distinct sequence allows the subsequent stoichiometric amplification of a plurality of templates of distinct sequence.01-01-2009
20090209008THERMUS THERMOPHILUS NUCLEIC ACID POLYMERASES - The invention provides novel nucleic acid polymerases from strains GK24 and RQ-1 of 08-20-2009
20090117621Methods of nucleic acid amplification and sequencing - The invention relates to a method of amplifying one or more nucleic acid templates on a solid support in a nucleic acid amplification reaction, for example by solid-phase PCR using one or more amplification primers attached to the solid support. The method is characterised in that the amplification primers used comprise a template-specific portion which is a sequence of at least 26 consecutive nucleotides and are not capable of annealing to target regions in the template under conditions of the amplification reaction. The method is particularly useful for amplifying human genomic DNA.05-07-2009
20090253183Amplicon Rescue Multiplex Polymerase Chain Reaction for Amplification of Multiple Targets - Disclosed is a method for amplifying and detecting polynucleotides which can provide sensitive, specific detection of multiple targets from a clinical specimen within a relatively short time.10-08-2009
20090275087ASYMMETRIC ADAPTER LIBRARY CONSTRUCTION - The present invention provides methods and compositions for asymmetrically tagging a nucleic acid fragment using asymmetric adapters.11-05-2009
20110229939METHOD FOR AMPLIFYING DOUBLE STRANDED TARGET SEQUENCE IN DOUBLE STRANDED DNA - The present invention relates to a nested PCR with high specificity. The present invention provides a method for amplifying a target sequence (09-22-2011
20120196330ISOTHERMAL DNA AMPLIFICATION - Provided herein are nucleic acid synthesis methods and agents that employ an endonuclease for example, endonuclease V, to introduce a nick into a target DNA including one or more inosine, and uses a DNA polymerase to generate amplicons of the target DNA.08-02-2012
20130122551Reducing Template Independent Primer Extension and Threshold Time for Loop Mediated Isothermal Amplification - Compositions and methods are provided for loop mediated isothermal amplification in which single stranded binding proteins are shown to protect primers from non-specific extension and to stimulate the rate of threshold amplification.05-16-2013
20130122552LOW-MASS SAMPLE BLOCK WITH RAPID RESPONSE TO TEMPERATURE CHANGE - A sample block for use in the polymerase chain reaction, DNA sequencing, and other procedures that involve the performance of simultaneous reactions in multiple samples with temperature control by heating or cooling elements contacting the bottom surface of the block is improved by the inclusion of hollows in the block that are positioned to decrease the mass of the block in the immediate vicinity of the wells.05-16-2013
20100159530COMPOSITIONS AND METHODS TO DETECT CANDIDA ALBICANS NUCLEIC ACID - Compositions are disclosed as nucleic acid sequences that may be used as amplification oligomers, including primers, capture probes for sample preparation, and detection probes specific for 06-24-2010
20080274514PENETRABLE CAP HAVING SPACED-APART GROOVES - A cap having a plurality of spaced-apart grooves in a downwardly tapered inner wall of the cap to facilitate the formation of air passageways as the cap is penetrated by a pipette tip. The air passageways aid in venting air from a fluid-holding vessel closed with the cap.11-06-2008
20110177563HIGH THROUGHPUT DEVICE FOR PERFORMING CONTINUOUS-FLOW REACTIONS - A high-throughput device is structured to perform a continuous-flow reaction, e.g., a polymerase chain reaction (PCR) requiring repetitive temperature control in a timely fashion.07-21-2011
20110177562Portable Preparation, Analysis, and Detection Apparatus for Nucleic Acid Processing - The present teachings comprise a device and method for lysing and/or purifying biological sample. The device can comprise a cartridge having a chamber containing a biological sample receiving region, a plurality of electrodes, and one or more sieving matrices. The electrodes can be configured to lyse the biological sample through the production of a pulsed electrical field. The electrodes can also be configured to heat lyse the biological sample. The electrodes can also be configured to electrophoretically move the biological sample through one or more sieving matrices. A portion of the sample can be isolated on a membrane. The portion of the sample isolated on the membrane can be amplified and detected. A portion of the sample can be isolated in a collection area present in the cartridge. The portion of the sample isolated in the collection area can be removed from the cartridge.07-21-2011
20100184155METHOD FOR REDUCING DISPERSION IN NUCLEIC ACID AMPLIFICATION REACTION - It is an object of the present invention to provide a method for amplifying a nucleic acid, which does not require complicated temperature control and which can be carried out without using special enzyme or special primers. The present invention provides a method for amplifying a nucleic acid, which comprises the following steps (1) and (2): (1) a step of incubating a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase, at least two types of oligonucleotide primers, and a nucleic acid fragment acting as a template, at temperature (T07-22-2010
20100184153High Multiplex Nucleic Acid Amplification - This invention relates to the amplification of multiple nucleic acid target sequences. Forward and reverse compound primers comprising a common amplification sequence and a target-specific primer sequence are immobilised at a site on a solid support. Target regions of single-stranded template DNA are primed and copied by the forward primer to produce a first extension product. The template DNA is removed and the first extension product is primed and copied by the reverse primer to produce a second extension product. The second extension product has common amplification sequences at each end and is bulk-amplified in solution by regular PCR employing primers that target the common amplification sequences. These methods allow highly multiplexed amplifications to be performed.07-22-2010
20100184154METHOD FOR REPLICATING NUCLEIC ACID SEQUENCE - It is an object of the present invention to provide a method for replicating a nucleic acid sequence using oligonucleotide primers and DNA polymerase. The present invention provides a method for replicating a nucleic acid sequence, which comprises synthesizing a complementary strand with a polymerase that catalyzes a strand displacement complementary strand synthesis reaction, wherein a double-stranded template nucleic acid having a sequence A(Ac) consisting of 20 or more to 200 or less contiguous nucleotides at both ends is used as an origin.07-22-2010
20100184152TARGET-ORIENTED WHOLE GENOME AMPLIFICATION OF NUCLEIC ACIDS - Disclosed herein are methods of amplifying target nucleic acid sequences (e.g., DNA or RNA), particularly from a very small amount of starting material, such as a single cell. These methods involve targeting the amplification of specific sequence(s) by use of sequence-specific primers and random primers for whole genome amplification using multiple displacement amplification. Generally, the provided methods are referred to herein as “target-oriented” whole genome amplification. Starting material for target-oriented whole genome amplification can be any sample containing DNA or RNA, however, the technique is particularly suitable for very small amounts of starting material, such as a few cells, a single cell, or a single nucleus. The methods provide amplified nucleic acid (including the target sequence of interest) that can subsequently be analyzed.07-22-2010
20100261230SYSTEM COMPRISING DUAL-SIDED THERMAL CYCLER AND EMULSION PCR IN POUCH - A system and method are provided for large volume sample amplification adaptable for use with conventional PCR-based reactions as well as emulsion-based PCR reactions. A sample is retained in a pouch or flexible bag which permits bulk PCR amplification with efficient heat-transfer properties. For applications involving emulsion-based PCR amplification, the system and method provide improved uniformity in emulsion amplification and can be used to amplify large or small volume emulsions rapidly and reproducibly.10-14-2010
20100216193Reaction chip, reaction method, temperature controlling unit for gene treating apparatus and gene treating apparatus - The reaction chip of the present invention has a plurality of recesses 08-26-2010
20100167356Homologous recombination-based DNA cloning methods and compositions - Methods and compositions for cloning a donor DNA molecule into an acceptor vector at a predetermined location are described. The methods are based on homologous recombination mediated by in vitro treatment of the donor DNA and the acceptor vector with an enzyme cocktail containing an exonuclease and a single-stranded DNA binding protein.07-01-2010
20100159529DRY COMPOSITION OF REACTION COMPOUNDS WITH STABILIZED POLMERASE - The present invention provides methods to obtain dry compositions of reaction compounds that maintain the biological activity of the compounds upon re-solubilization after a certain storage time. Preferably, the dry composition comprises a polymerase, and the dry composition is usable for polymerase chain reaction (PCR) amplification after re-solubilization.06-24-2010
20110059490POLYMERASE COMPOSITIONS AND USES - A composition having nucleic acid polymerase activity, which comprises an active nucleic acid polymerase and an excess amount of a non-functional mutant nucleic acid polymerase protein, wherein the non-functional mutant nucleic acid polymerase protein stabilizes the active nucleic acid polymerase against loss of polymerase activity.03-10-2011
20100255546NUCLEIC ACID AMPLIFICATION METHOD - Disclosed is a novel isothermal nucleic acid amplification method enabling inexpensive and simple and easy detection. The method includes introducing nicking enzyme recognition sequences into an analysis target nucleic acid using nicking enzyme recognition sequence-containing primers and isothermally amplifying a specific region of the target nucleic acid using the primers, a nicking enzyme and a DNA polymerase having strand displacement activity.10-07-2010
20100233763DUAL-SIDED THERMAL CYCLER - A device, system, and method are provided for thermally treating a fluid processing device. According to various embodiments, a system is provided that can include a thermal device and a fluid processing device holder. The thermal device can include a first block having a thermal conductivity greater than 0.5 Watt per centimeter Kelvin (W/cm·K), a second block having a thermal conductivity greater than 0.5 W/cm·K, and a heat-pump device disposed between the first block and the second block. The heat-pump device can transfer thermal energy from at least one of the first block and the second block to the other of the first block and the second block. The fluid processing device holder can hold a fluid processing device in a heat-transfer position with respect to the first block and the second block. The fluid processing device can be a microfluidic device.09-16-2010
20100240103OLIGONUCLEOTIDES LABELED WITH A PLURALITY OF FLUOROPHORES - An embodiment of the invention discloses new methods for designing labeled nucleic acid probes and primers by labeling oligonucleotides with a plurality of spectrally identical or similar dyes and optionally with one or more quencher dyes. Oligonucleotides labeled in accordance with some embodiments of the invention exhibit a detectable increase in signal, for example, fluorescent signal when the labeling dyes are separated from one another. Methods for separating the dye include cleaving the labeled oligonucleotides include using enzymes that have 5′-exonuclease activity. In one embodiment nucleic acid primers of the present invention may fluoresce upon hybridization to a target sequence and incorporation into the amplification product. Nucleic acid probes and primers of the present invention have wide applications ranging from general detection of a target nucleic acid sequence to clinical diagnostics. Major advantages of the oligonucleotides including nucleic acid probes and primers of many embodiments of the present invention are their synthetic simplicity, spectral versatility and superior fluorescent signal.09-23-2010
20100129874METHOD FOR MULTIPLEXED NUCLEIC ACID PATCH POLYMERASE CHAIN REACTION - The invention encompasses a method for amplifying at least two different nucleic acid sequences. In particular, the method encompasses a multiplexed nucleic acid patch polymerase chain reaction.05-27-2010
20130217071METHODS AND COMPOSITIONS FOR PERFORMING NUCLEIC ACID AMPLIFICATION REACTIONS - The present disclosure provides methods and compositions for performing nucleic acid reactions, such as the reverse transcriptase-polymerase chain reaction (RT-PCR).08-22-2013
20100255547POLYANION FOR IMPROVED NUCLEIC ACID AMPLIFICATION - The present invention is directed to a novel chemical compound comprising the structure [Xx-(CH2)m-phosphate-Yy]n, characterized in that 3≦m≦6, 30≦n≦60, each x and y is independently from each other 0 or 1, each X and Y is independently from each other any photometrically measurable entity; provided that the terminal X can also be an —OH group or a phosphate group, and further provided that the terminal Y can also be an —OH group. Such a compound can be used as a suitable hot start additive for PCR based amplification of nucleic acids.10-07-2010
20090042256REACTOR PLATE AND REACTION PROCESSING METHOD - Disclosed herein is a reactor plate including a sealed reaction well, a reaction well channel connected to the reaction well, and a reaction well air vent channel connected to the reaction well. The reaction well contains a reagent contained in the bottom thereof and a thermally fusible material which is solid at room temperature and is contained in the bottom thereof to encapsulate the reagent. When the reaction well is heated, the thermally fusible material is melted, and as a result, a sample liquid introduced into the reaction well through the reaction well channel reacts with the reagent in the reaction well.02-12-2009
20120196329POLYMERIZATION OF NUCLEIC ACIDS USING ACTIVATION BY POLYPHOSPHOROLYSIS (APP) REACTIONS - This disclosure relates to methods of performing activation by polyphosphorolysis (APP) reactions using at least one of the polyphosphorylating agents triphosphate, polyphosphate, imidodiphosphate, thiodiphosphate (or μ-monothiopyrophosphate), and related compounds.08-02-2012
20090075345Methods for Genotyping with Selective Adaptor Ligation - The present invention provides methods for reducing the complexity of a nucleic acid sample to interrogate a collection of target sequences. Complexity reduction can be accomplished by fragmenting the nucleic acid sample with a restriction enzyme that has at least one variable position in the recognition sequence. In some aspects adaptors that ligate to some but not all possible overhangs generated by digestion are ligated to the fragments. This selective adaptor ligation allows for selective amplification of a subset of the fragments using primers complementary to the adaptor sequence. In another aspect primers that are complementary to a subset of the fragments after adaptor ligation are used for amplification. Amplified fragments may be analyzed to genotype polymorphisms by hybridization to an array of probes that are complementary to target sequences that will be amplified.03-19-2009
20090075344SAMPLE PREPARATION APPARATUS - A reconfigurable sample preparation device includes a rotary plunger device having a hollow body and a coupling device, provided above one end of the rotary plunger, and accommodating a sample. The device also includes at least one sealed reagent module. When the rotary plunger is rotated on the coupling device, a film of the reagent module is pierced, mixing the sample with a substance from the reagent module.03-19-2009
20090075343SELECTION OF DNA ADAPTOR ORIENTATION BY NICKING - Aspects described and claimed herein provide methods to insert multiple DNA adaptors into a population of circular target DNAs at defined positions and orientations with respect to one another using nicking reactions. The resulting multi-adaptor constructs are then used in massively-parallel nucleic acid sequencing techniques.03-19-2009
20120142060Compositions and Methods for Processing and Amplification of DNA, Including Using Multiple Enzymes in a Single Reaction - The present invention concerns preparation of DNA molecules, such as a library, using a stem-loop oligonucleotide. In particular embodiments, the invention employs a single reaction mixture and conditions. In particular, at least part of the inverted palindrome is removed during the preparation of the molecules to facilitate amplification of the molecules. Thus, in specific embodiments, the DNA molecules are suitable for amplification and are not hindered by the presence of the palindrome.06-07-2012
20110027832USE OF TAQ POLYMERASE MUTANT ENZYMES FOR NUCLEIC ACID AMPLIFICATION IN THE PRESENCE OF PCR INHIBITORS - The present invention generally relates to detection of a target nucleic acid in standard PCR, real-time PCR, RT PCR, and real-time RT PCR. One aspect of the invention provides mutant DNA polymerase enzymes that are resistant to PCR inhibitors, such as dye, blood, and soil. Another aspect of the invention provides for methods of real-time PCR assays using mutant DNA polymerase enzymes resistant to PCR inhibitors with samples containing dye, blood, and/or soil. Another aspect of the invention provides for methods of standard PCR assays using mutant DNA polymerase enzymes resistant to PCR inhibitors with samples containing blood and/or soil.02-03-2011
20090170168METHOD FOR MAKING AVAILABLE A PRIMING OLIGONUCLEOTIDE - The present invention is directed to novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic (i.e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, the present invention discloses a method of nucleic acid amplification which is robust and efficient, while reducing the appearance of side-products. The method uses only one primer, the “priming oligonucleotide,” a promoter oligonucleotide modified to prevent polymerase extension from its 3′-terminus and, optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or substantially eliminating the formation of side-products. The method of the present invention minimizes or substantially eliminates the emergence of side-products, thus providing a high level of specificity. Furthermore, the appearance of side-products can complicate the analysis of the amplification reaction by various molecular detection techniques. The present invention minimizes or substantially eliminates this problem, thus providing an enhanced level of sensitivity.07-02-2009
20090111149METHOD OF REDUCING CROSS-CONTAMINATION IN CONTINUOUS AMPLIFICATION REACTIONS IN A CHANNEL - The present invention relates to a method for reducing cross-contamination in continuous amplification reactions in channels of microfluidic devices. More specifically, the present invention relates to the use of specific materials continuously flowing in the channels to reduce adsorption of MgCl04-30-2009
20100221787ISOTHERMAL AMPLIFICATION METHOD AND DNA POLYMERASE USED IN THE SAME - A DNA polymerase suitable for specific isothermal amplification methods and an isothermal amplification method using the DNA polymerase are provided. In the presence of a DNA polymerase including a protein described in the following item (a) or (b), an amplification reaction of a target nucleic acid sequence in a nucleic acid sample is carried out isothermally using a first primer shown in the following (X). By using the DNA polymerase, it becomes possible to carry out the amplification reaction using the primer within a shorter time than ever before. 09-02-2010
20100184156Recombinant Colwellia Psychrerythraea Alkaline Phosphatase and Uses Thereof - A heat labile alkaline phosphatase enzyme and methods of using the same and kits including the same are disclosed. Specifically, a nucleotide sequence of, peptide sequence of, methods of using, and kits comprising, a heat labile alkaline phosphatase isolated from 07-22-2010
20100297707REACTION VESSEL COMPRISING CONDUCTIVE LAYER AND INNER NON-METALLIC LAYER - A reaction vessel for conducting a chemical or biochemical reaction, such as a polymerase chain reaction wherein at least one wall of said vessel comprises a metallic layer and an inner non-metallic layer. Reaction systems comprising combinations of vessels of the invention and apparatus for heating them, as well as particular reactions vessels are also described and claimed.11-25-2010
20100297708MOLECULAR DIAGNOSTICS SYSTEM AND METHODS - The present invention relates to automated devices and methods for the extraction of nucleic acids from cells, the amplification of segments of nucleic acid and the detection of nucleic acids, all in a convenient and portable manner. The invention is particularly suited for use in point-of-care medical diagnostics testing.11-25-2010
20100304442METHODS, COMPOSITIONS AND KITS FOR THE IMPROVED DETECTION OF SMALL RNA MOLECULES - The present invention provides compositions, methods and kits for use in the detection of small RNA sequences, which allow for rapid and robust amplification and detection. The methods provide improved sensitivity and efficiency in the amplification-based detection of small RNA sequences by incorporating one or more base-modified duplex-stabilizing dNTPs during reverse transcription and/or amplification.12-02-2010
20110033899CONVECTION POLYMERASE CHAIN REACTION METHOD - The invention relates to a method for rapid detection of specific nucleic acid fragments with the aid of a PCR, consisting in amplifying target products in a special DNA thermocycler, provided with a special reaction thermal unit, which makes it possible to obtain, in reaction vessels in the form of standard polypropylene test tubes, a sloping temperature gradient directed at an angle to the direction of gravity. The amplification time is of 1-5 minutes. The inventive method can be recommended for DNA diagnostics, also field conditions, in medicine, veterinary sciences, in sanitary and epidemiological studies for detecting agents of dangerous infections, including potential bio-terrorist attacks, in criminalistics for identifying criminals in the food industry for detecting food products from genetically modified organisms, for testing raw material quality etc.02-10-2011
20090068708Method for molecular cloning and polynucleotide synthesis using vaccinia DNA topoisomerase - This invention provides a modified vaccinia topoisomerase enzyme containing an affinity tag which is capable of facilitating purification of protein-DNA complexes away from unbound DNA. This invention further provides a modified sequence specific topoisomerase enzyme.03-12-2009
20110117608DOUBLE-STRANDED NUCLEIC ACID - The invention is directed towards constructs for RNAi techniques. The invention provides a ribonucleic acid (RNA) for use as interfering RNA in gene silencing techniques to silence a target gene comprising in a 5′ to 3′ direction at least a first effector sequence, a second effector sequence, a sequence substantially complementary to the second effector sequence and a sequence substantially complementary to the first effector sequence, wherein the complementary sequences are capable of forming double stranded regions with their respective effector sequences and wherein at least one of these sequences is substantially identical to the predicted transcript of a region of the target gene, and a nucleic acid construct encoding such an RNA.05-19-2011
20110008846MODIFIED ENZYMES AND THEIR USES - The invention relates to an active enzyme of bacterial, fungal, viral, or archae origin, wherein the enzyme is coupled, preferably covalently coupled with at least one polymer having a molecular weight between about 500 to about 20,000 daltons selected from the group consisting of polyethylene glycol and polypropylene glycol. The invention relates in particular to uses thereof of such as in molecular biology as well as kits comprising such enzymes. In preferred embodiments the enzymes of the invention are nucleic acid modifying or replicating enzymes.01-13-2011
20110008848ENZYME - There is provided a polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 55% identity to 01-13-2011
20110008847METHOD OF ISOLATING NUCLEIC ACIDS FROM A BIOLOGICAL SAMPLE - Method of preparing a biological sample appropriate for use in a subsequent in vitro nucleic acid amplification reaction. The method involves a rapid, transient exposure to alkaline conditions which can be achieved by mixing an alkaline solution with a pH-buffered solution that includes a detergent and the biological sample to be tested for the presence of particular nucleic acid species using in vitro amplification. The invented method advantageously can improve detection of some target nucleic acids without substantially compromising detectability of others. The method is particularly useful for simultaneously preparing RNA and DNA templates that can be used in multiplex amplification reactions.01-13-2011
20130149749POLYMERASE - An engineered DNA polymerase characterised in that the polymerase exhibits an enhanced ability to process nucleic acid in the presence of environmental and biological inhibitors compared to wild type DNA polymerase.06-13-2013
20110039303MICROFLUIDIC AND NANOFLUIDIC DEVICES, SYSTEMS, AND APPLICATIONS - The present invention discloses the integration of programmable microfluidic circuits to achieve practical applications to process biochemical and chemical reactions and to integrate these reactions. In some embodiments workflows for biochemical reactions or chemical workflows are combined. Microvalves such as programmable microfluidic circuit with Y valves and flow through valves are disclosed. In some embodiments microvalves of the present invention are used for mixing fluids, which may be part of an integrated process. These processes include mixing samples and moving reactions to an edge or reservoir for modular microfluidics, use of capture regions, and injection into analytical devices on separate devices. In some embodiments star and nested star designs, or bead capture by change of cross sectional area of a channel in a microvalve are used. Movement of samples between temperature zones are further disclosed using fixed temperature and movement of the samples by micropumps.02-17-2011
20110117609Homologous Recombination Method, Cloning Method, and Kit - Disclosed is a method which can achieve the homologous recombination of a gene of interest selectively. Also disclosed is a recombinant DNA molecule produced by the method. Specifically disclosed is a homologous recombination method which uses a PCR product and a linearized vector. The PCR product comprises a sequence for a target gene and amplification primer sequences P05-19-2011
20110111462Composition and Method for Synthesizing a Deoxyribonucleotide Chain Using a Double Stranded Nucleic Acid Complex with a Thermostable Polymerase - The present invention relates to the field of molecular biology, and more particular, to a nucleic acid construct for use in amplification processes. More precisely, the invention enhances the specificity of amplification of nucleic acids by means of a double stranded oligonucleotide modified with a molecule having the ability to prevent extension of the double stranded nucleic acid.05-12-2011
20110117607ANNULAR COMPRESSION SYSTEMS AND METHODS FOR SAMPLE PROCESSING DEVICES - Systems and methods for processing sample processing devices. The system can include a base plate adapted to rotate about a rotation axis. The base plate can include at least one first magnetic element. The system can further include an annular cover, and a sample processing device comprising at least one thermal process chamber. The annular cover can include an inner edge, an outer edge, and at least one second magnetic element. The method can include positioning the sample processing device between the base plate and the annular cover, such that the inner edge of the annular cover is positioned inwardly of the at least one thermal process chamber, and such that the at least one first magnetic element attracts the at least one second magnetic element to force the annular cover in a first direction along the z-axis, urging the sample processing device into contact with the base plate.05-19-2011
20110212492PCR METHOD AND PCR DEVICE - Provided on an inner surface of a container to carry out polymerase chain reaction (PCR) are an electrode pair 09-01-2011
20110086393METHOD TO PRODUCE SINGLE STRANDED DNA OF DEFINED LENGTH AND SEQUENCE AND DNA PROBES PRODUCED THEREBY - A method for producing a single stranded DNA (ssDNA) molecule of a defined length and sequence is disclosed. This method enables the preparation of, inter alia, probes of greater length than can be chemically synthesized. The method starts with a double stranded molecule, such as genomic, double stranded DNA (dsDNA) from any organism. A fragment of the starting molecule (dsDNA) is amplified by specific primers engineered to introduce cleavage sites on either side of the desired sequence. Cleavage steps on the amplified, engineered fragment are combined with a phosphate removal step, thereby creating a construct that can be digested with an exonuclease without damage to the desired ssDNA. Probes, which hybridize with large gaps between the ends of the probes, are also disclosed.04-14-2011
20110244521METHOD FOR PREPARATION OF RECOBINANT DNA - The problem to be solved in the present invention is to provide a simplified and efficiently improved DNA recombination method.10-06-2011
20110124052Multiplex Targeted Amplification Using Flap Nuclease - Methods for multiplex amplification of a plurality of targets of distinct sequence from a complex mixture are disclosed. In one aspect targets are circularized using a single circularization probe that is complementary to two regions in the target that flank a region to be amplified. The targets may hybridize to the circularization probe so that 5′ or 3′ flaps are generated and methods for removing flaps and circularizing the resulting product are disclosed. In another aspect targets are hybridized to dU probes so that 5′ and 3′ flaps are generated. The flaps are cleaved using 5′ or 3′ flap endonucleases or 3′ to 5′ exonucleases. The target sequences are then ligated to common primers, the dU probes digested and the ligated targets amplified.05-26-2011
20110124051PYROPHOSPHOROLYSIS ACTIVATED POLYMERIZATION (PAP) - A novel method of pyrophosphorolysis activated polymerization (PAP) has been developed. In PAP, pyrophosphorolysis and polymerization by DNA polymerase are coupled serially for each amplification by using an activatable oligonucleotide P* that has a non-extendible 3′-deoxynucleotide at its 3′ terminus. PAP can be applied for exponential amplification or for linear amplification. PAP can be applied to amplification of a rare allele in admixture with one or more wild-type alleles by using an activatable oligonucleotide P* that is an exact match at its 3′ end for the rare allele but has a mismatch at or near its 3′ terminus for the wild-type allele. PAP is inhibited by a mismatch in the 3′ specific sequence as far as 16 nucleotides away from the 3′ terminus. PAP can greatly increase the specificity of detection of an extremely rare mutant allele in the presence of the wild-type allele. Specificity results from both pyrophosphorolysis and polymerization since significant nonspecific amplification requires the combination of mismatch pyrophosphorolysis and misincorporation by the DNA polymerase, an extremely rare event. Using genetically engineered DNA polymerases greatly improves the efficiency of PAP.05-26-2011
20110244522ROTATIONAL PCR EQUIPMENT AND PCR METHOD USING THE SAME - Provided are a rotational PCR apparatus, a PCR chip for the same and a rotational PCR method using the same.10-06-2011
20090246834Nano-PCR: methods and devices for nucleic acid amplification and detection - Methods, devices, and compositions are described that provide for amplification of nucleic acid sequences without reliance upon temperature cycling, thus freeing the methods from conventional benchtop thermal cycling devices. Denaturation of double stranded nucleic acids, primer annealing, and precision control over primer extension by polymerase can be accomplished by applying stress to a nucleic acid. These methods can provide one or more benefits over conventional PCR methods including: precision control over the PCR process; generally improved fidelity; improved accuracy over problematic sequences such as GC-rich or tandem repeat regions; greater sequence length; increased reaction yield; reduced experimental time; greater efficiency; lower cost; greater portability; and, robustness to various environmental parameters, such as temperature, pH, and ionic strengths.10-01-2009
20090325235THERMOACTIVE SIVagm SAB REVERSE TRANSCRIPTASE - Methods and kits performing reverse transcription and RT-PCR reactions having high fidelity, processivity and DNA polymerase activity are described. The methods involve performance of reverse transcription at an increased temperature with a reverse transcriptase from Simian Immunodeficiency Virus-agm.sab or a variation thereof. The kits of the present invention include a reverse transcriptase from Simian Immunodeficiency Virus-agm.sab or a variation thereof, a DNA polymerase capable of amplifying cDNA under conditions suitable for polymerase chain reaction, and the reagents necessary to carry out both processes.12-31-2009
20100015668Programmable Oligonucleotide Synthesis - The invention relates to methods and devices for preparing synthetic nucleic acids.01-21-2010
20100055743LOW-MASS SAMPLE BLOCK WITH RAPID RESPONSE TO TEMPERATURE CHANGE - A sample block for use in the polymerase chain reaction, DNA sequencing, and other procedures that involve the performance of simultaneous reactions in multiple samples with temperature control by heating or cooling elements contacting the bottom surface of the block is improved by the inclusion of hollows in the block that are positioned to decrease the mass of the block in the immediate vicinity of the wells.03-04-2010
20100055742METHOD FOR AMPLIFICATION OF NUCLEOTIDE SEQUENCE - To provide a method for electively amplifying a target nucleic acid and a method for detecting a nucleic acid by said method, which are useful as a method for synthesizing the nucleic acid. A method for amplifying a nucleic acid sequence [EVA (Endonuclease V-dependent Amplification) method] which selectively amplifies a target nucleic acid in a sample, by the use of an oligonucleotide primer containing a base which can be recognized by an endonuclease V, the endonuclease V and a DNA polymerase having a strand displacement activity, and a method for detecting a nucleic acid by said method.03-04-2010
20110097762LINKERS AND CO-COUPLING AGENTS FOR OPTIMIZATION OF OLIGONUCLEOTIDE SYNTHESIS AND PURIFICATION ON SOLID SUPPORTS - A method of modulation of synthesis capacity on and cleavage properties of synthetic oligomers from solid support is described. The method utilizes linker molecules attached to a solid surface and co-coupling agents that have similar reactivities to the coupling compounds with the surface functional groups. The preferred linker molecules provide an increased density of polymers and more resistance to cleavage from the support surface. The method is particularly useful for synthesis of oligonucleotides, oligonucleotides microarrays, peptides, and peptide microarrays. The stable linkers are also coupled to anchor molecules for synthesis of DNA oligonucleotides using on support purification, eliminating time-consuming chromatography and metal cation presence. Oligonucleotides thus obtained can be directly used for mass analysis, DNA amplification and ligation, hybridization, and many other applications.04-28-2011
20100062494ENZYMATIC OLIGONUCLEOTIDE PRE-ADENYLATION - Methods and compositions for making and using pre-adenylated oligonucleotide sequences are provided.03-11-2010
20100068765SYSTEMS AND METHODS FOR THE AMPLIFICATION OF DNA - A system for amplifying nucleic acids is disclosed which, in one embodiment, includes a fluidic device having a sample channel and a heat exchange channel disposed sufficiently close to the sample channel such that a heat exchange fluid in the heat exchange channel can cause a sample in the sample channel to gain or lose heat at desired levels. In one illustrative embodiment, the system further includes three reservoirs coupled to the heat exchange channel and a temperature control system configured to heat fluids stored in the respective reservoirs at different temperatures. One or more pumps and a controller are configured to cause fluid stored in the reservoirs to enter and flow through the heat exchange channel at different times.03-18-2010
20110081685COMPOSITIONS AND METHODS FOR PROCESSING AND AMPLIFICATION OF DNA, INCLUDING USING MULTIPLE ENZYMES IN A SINGLE REACTION - The present invention concerns preparation of DNA molecules, such as a library, using a stem-loop oligonucleotide. In particular embodiments, the invention employs a single reaction mixture and conditions. In particular, at least part of the inverted palindrome is removed during the preparation of the molecules to facilitate amplification of the molecules. Thus, in specific embodiments, the DNA molecules are suitable for amplification and are not hindered by the presence of the palindrome.04-07-2011
20090215126NUCLEIC ACID-FREE THERMOSTABLE ENZYMES AND METHODS OF PRODUCTION THEREOF - The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).08-27-2009
20090215125DEVICES AND PROCESSES FOR NUCLEIC ACID EXTRACTION - Devices, processes, and kits for the extraction of nucleic acids from biological samples are disclosed. The devices comprise a first port, a second port, and a binding chamber intermediate and in fluid communication with the first port and the second port. The binding chamber comprises an unmodified flat glass surface effective for binding a heterogeneous population of nucleic acids. The first port, second port, and binding chamber define a continuous fluid pathway that is essentially free of nucleic acid-specific binding sites.08-27-2009
20090215124Nucleic acid isolation methods and materials and devices thereof - The present invention relates to methods for purifying nucleic acid from a sample using mild conditions that do not affect the chemical integrity of the nucleic acid. The method comprises contacting the sample with an matrix entrapped chitosan solid phase which is able to bind the nucleic acids at a first pH, and then extracting the nucleic acid from the solid phase by using an elution solvent at a second pH.08-27-2009
20110097763Thermal Cycling Method - The invention relates to certain novel approaches to reducing or eliminating the movement of contaminants from one droplet to another on a droplet actuator via liquid filler fluid. In one application, droplet actuators are used to conduct genetic analysis using polymerase chain reaction (PCR) techniques. The invention addresses the need for improved methods of performing PCR on a droplet actuator that provide for optimum amplification and detection of a sample target.04-28-2011
20110081687Enrichment Through Heteroduplexed Molecules - The present invention relates to the enrichment of specific target sequences. Enrichment can be achieved through the formation of a heteroduplex that includes the specific target sequence and then the specific cleavage of the heteroduplex. A binding moiety is then added to the cleaved heteroduplex, allowing for the subsequent manipulation of the specific target sequence in the heteroduplex.04-07-2011
20110081686Reactor for Bulk Manufacture of PCR Amplicons - A reactor and methods of use are disclosed for amplification of an amplicon. The reactor includes a housing having a fluid path with a fluid inlet and a fluid outlet and one or more reaction chambers suitable for housing the amplicon. The reaction chambers are positioned within the housing in thermal communication with a fluid flowing in the fluid path to heat and/or cool the reaction chambers and the inside of the reaction chambers are coated with or are a plastic material. A variable temperature fluid source is in fluid communication with the fluid inlet and the fluid source is configured to provide fluid at temperatures suitable for amplification, including a denature temperature, an anneal temperature, and an extension temperature. A controller is coupled to the variable temperature fluid source to control a fluid temperature within the fluid path by controlling the variable temperature fluid source.04-07-2011
20110076727PENETRABLE CAP - A cap having a core structure dimensioned to receive a pipette therethrough. The cap includes two axially aligned frangible seals that are affixed to the core structure in a spaced-apart relationship. The frangible seals are constructed so that air passageways are formed between the frangible seals and a pipette tip when the pipette tip penetrates the frangible seals. The cap optionally includes a filter interposed between the first and second frangible seals.03-31-2011
20100279359TEMPLATE-DEPENDENT NUCLEIC ACID POLYMERIZATION USING OLIGONUCLEOTIDE TRIPHOSPHATES BUILDING BLOCKS - A novel use of a template-dependent polymerase. The novel use is effected by employing the template-dependent polymerase for incorporating at least one oligonucleotide triphosphate onto a nascent oligonucleotide-3′-OH in a template-dependent manner.11-04-2010
20100311126METHOD FOR IN VITRO RECOMBINATION - The present invention relates, e.g., to an in vitro method, using isolated protein reagents, for joining two double-stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising (a) chewing back the DNA molecules with an enzyme having an exonuclease activity, to yield single-stranded overhanging portions of each DNA molecule which contain a sufficient length of the region of sequence identity to hybridize specifically to each other; (b) specifically annealing the single-stranded overhangs; and (c) repairing single-stranded gaps in the annealed DNA molecules and sealing the nicks thus formed (ligating the nicked DNA molecules). The region of sequence identity generally comprises at least 20 non-palindromic nucleotides (nt), e.g., at least about 40 non-palindromic nt. In some embodiments of the invention, about 5% PEG is present during all steps of the reaction, and/or the repair reaction is achieved with Taq DNA polymerase and a compatible ligase, such as Taq DNA ligase. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes. It can be used, e.g., to join synthetically produced sub-fragments of a gene or genome of interest.12-09-2010
20100304444METHOD OF AMPLIFICATION - The present invention relates generally to a method of amplifying a nucleic acid region of interest and, more particularly, to a method of amplifying a nucleic acid region of interest via a nested single tube PCR. The method of the invention is designed to provide a means to selectively inactivate the functionality of the outer primer or primers and to maintain amplification efficiency throughout the reaction. The development of a means to achieve efficient amplification by the outer primer followed by efficient amplification with the inner primers, in the context of a single tube nested PCR, is useful in a range of applications including, but not limited to, the diagnosis and/or monitoring of disease conditions which are characterized by specific gene sequences and the characterization or analysis of specific gene regions of interest. Still further, the method of the present invention enables quantification to be performed and not just simple detection.12-02-2010
20110250649PCR-BASED METHOD OF SYNTHESIZING A NUCLEIC ACID MOLECULE - There is provided a method of synthesizing a nucleic acid molecule and in one aspect, the method comprises assembling a full length template nucleic acid molecule by PCR in a PCR reaction mixture comprising a set of assembly oligonucleotides having a first average melting temperature and a set of outer amplification primers having a second average melting temperature that is lower than the first average melting temperature, wherein said assembling comprises subjecting the PCR reaction mixture to a first annealing temperature that is higher than the second average melting temperature and; amplifying the full length template nucleic acid molecule by PCR in the PCR reaction mixture wherein said amplifying comprises subjecting the PCR reaction mixture to a second annealing temperature that permits annealing of the outer amplification primers to the full length template nucleic acid molecule.10-13-2011
20110250648Compositions And Methods For Inhibiting Terminal Transferase Activity - The present invention relates to systems and methods for amplifying nucleic acid. In particular, systems and methods are provided for inhibiting polymerase based terminal transferase activity within a polynucleotide amplification setting (e.g., polymerase chain reaction). In addition, systems and methods are provided for generating amplified products generated with polynucleotide amplification techniques having reduced 3′ non-templated nucleotide addition.10-13-2011
20110212490METHODS AND COMPOSITIONS FOR PERFORMING LOW BACKGROUND MULTIPLEX NUCLEIC ACID AMPLIFICATION REACTIONS - Methods and compositions for performing low background multiplex nucleic acid amplification reactions are provided. Aspects of the invention include contacting a nucleic acid sample with two or more primer pairs for two or more target nucleic acids under template dependent primer extension reaction conditions, e.g., polymerase chain reaction (PCR) conditions. The resultant amplified composition is then contacted with target nucleic acid circularizing reagents, and product circularized target nucleic acids are then selected, e.g., for further amplification. Also provided are systems and kits that find use in practicing embodiments of the inventions.09-01-2011
20100003725LOW-MASS SAMPLE BLOCK WITH RAPID RESPONSE TO TEMPERATURE CHANGE - A sample block for use in the polymerase chain reaction, DNA sequencing, and other procedures that involve the performance of simultaneous reactions in multiple samples with temperature control by heating or cooling elements contacting the bottom surface of the block is improved by the inclusion of hollows in the block that are positioned to decrease the mass of the block in the immediate vicinity of the wells.01-07-2010
20110151519Laboratory Apparatus with an Arrangement for the Tempering of Samples and Method of Tempering Samples - The invention relates to a laboratory apparatus, in particular for performing a polymerase chain reaction (PCR) in a plurality of PCR-samples, which comprises an arrangement for tempering samples, the arrangement comprising a tempering block for the tempering of samples, the tempering block comprising a reception side, which provides receptacles for receiving sample vessels, and a contact side for the contact of at least one tempering device, at least one tempering device, arranged in an area of said contact side, a pressure device, which comprises a pressure element and an auxiliary element, said at least one tempering device being arranged between said auxiliary element and the tempering block, the pressure element being linked to said auxiliary element and to the tempering block, and being arranged to press said at least one tempering device against the tempering block by pressing said auxiliary element against said at least one tempering device, wherein at least one tempering device is shaped and arranged in said area to at least partially surround by itself said pressure element.06-23-2011
20090317874SYSTEMS AND METHODS FOR AMPLIFYING NUCLEIC ACIDS - An apparatus for performing a thermocyclic process, such as amplifying DNA, includes a microfluidic chip with a channel formed therein and one or more thermal distribution elements disposed over portions of the chip. Each thermal distribution element is configured to distribute thermal energy from an external thermal energy source substantially uniformly over the portion of the chip covered by the thermal distribution element. The portion of the chip covered by the thermal distribution element thereby comprises a discrete temperature zone. Other temperature zones can be defined by other thermal distribution elements or by portions of the chip not covered by a thermal distribution element. The channel is configured so that a fluid flowing through the channel would enter and exit the different temperature zones a plurality of times, thereby alternately exposing the fluid to the temperature of each zone for a period of time required for the fluid to traverse the zone.12-24-2009
20080311628Methods and compositions for rapid amplification and capture of nucleic acid sequences - A method for amplifying a nucleic acid sequence includes the steps of (i) providing a first pair of primers that include one or more uracil nucleotides, the primers being complementary to a portion of a genomic template, (ii) introducing the first pair of primers, the genomic template and a first polymerase into a reaction vessel, (iii) carrying out one or more polymerase chain reaction cycles in the reaction vessel to generate a plurality of first amplicons, and (iv) selectively degrading a portion each first amplicon with a Uracil-DNA Glycosylase to decrease the binding energy of each first amplicon. In one embodiment, the step of selectively degrading includes using a thermostable Uracil-DNA Glycosylase to decrease the binding energy of each first amplicon. In another embodiment, the method also includes the step of adding a second polymerase and a second pair of primers to the reaction vessel to generate a plurality of second amplicons that are different than the first amplicons. Generating the plurality of second amplicons can occur substantially isothermally or non-isothermally. Further, the second pair of primers can be nested primers.12-18-2008
20110256591INDIVIDUALLY SYNTHESIZED PRIMERS TO BE USED IN WHOLE GENOME AMPLIFICATION - Disclosed are compositions and methods for random amplification of nucleic acid sequences of interest using random-G-deficient primers. Also disclosed are methods of randomly amplifying a target nucleic acid sequence using random G-deficient primers alone or in combination with random, partially random, or specific primers.10-20-2011
20080268507Recombinant Dna Nicking Endonuclease and Uses Thereof - Recombinant nicking endonucleases and associated methylases have been obtained and sequenced and their specificity has been defined. A mutant form of the nicking endonuclease has been cloned where the mutation includes deletion of amino acid sequences at the C-terminal end of the protein. The nicking enzymes have been used for a number of purposes including: amplifying DNA from as few cells as can be found in a single bacterial colony in the presence of a strand displacing polymerase; and for removing genomic DNA in a biological preparation where it is deemed to be a contaminant.10-30-2008
20100297706MUTANT DNA POLYMERASES AND THEIR GENES FROM THERMOCOCCUS - The present invention relates to mutant DNA polymerases and their genes isolated from 11-25-2010
20080254517Thermal Cycler With Optimized Sample Holder Geometry - The invention concerns a thermal cycler and a microtiter plate. The cycler comprises a sample holder having a first surface and a surface and means for automated, controlled heating and cooling of the sample holder. The first surface of the sample holder is designed to hold a plurality of samples arrayed in a grid having a predefined pitch. The number of samples in one dimension is an exact match of the SBS plate standards for that sample pitch and in another dimension corresponds to a fraction of the number of samples in a second dimension of an SBS microtiter standard plate. According to the invention, the sample holder is shaped such that the area of the second surface is larger than the area of the first surface. By means of the invention, the thermal ramping speeds of the cycler can be significantly increased.10-16-2008
20080254516Method of Isolating Nucleic Acid Targets - The invention provides efficient methods of isolating specific nucleic acid targets to obtain information from target nucleic acid sequences in a relatively short time period. DNA or cDNA is enzymatically digested into smaller fragments, double-stranded DNA linkers are added onto the ends of the DNA fragments to flank each fragment with a known DNA sequence. The fragments are mixed with an oligonucleotide probe that is bound to a marker and contains a conserved nucleic acid sequence of interest. The fragments that hybridize to the probe through nucleotide base pair complementation become indirectly connected to the marker. These target fragments are captured using a capture agent that specifically recognizes the marker and treated to prevent non-specific binding. Captured fragments are typically cloned prior to sequencing. The captured fragments may also be amplified using PCR to increase the efficiency of the cloning.10-16-2008
20110256590BIOCHIP, REACTOR, AND REACTION METHOD - A biochip includes: a chamber that has a longitudinal direction; a holding unit that holds a liquid sample within a predetermined area of the chamber provided along the longitudinal direction, and releases the liquid sample from the predetermined area to an area inside the chamber by using a predetermined pressing force; and a pressed member that applies the predetermined pressing force to the liquid sample.10-20-2011
20110027833THERMOSTABLE TYPE-A DNA POLYMERASE MUTANTS WITH INCREASED POLYMERIZATION RATE AND RESISTANCE TO INHIBITORS - The present invention provides mutants of DNA polymerases having an increased rate of incorporation of nucleotides into nucleic acids undergoing polymerization and having an enhanced resistance to inhibitors of DNA polymerase activity. The mutant polymerases are well suited for fast PCR applications, for PCR amplification of targets in samples that contain inhibitors of wild-type polymerases, and for fast PCR amplification of samples containing DNA polymerase inhibitors. In exemplary embodiments, the mutants are mutants of Taq DNA polymerase.02-03-2011
20100285537SELECTIVE TAGGING OF SHORT NUCLEIC ACID FRAGMENTS AND SELECTIVE PROTECTION OF TARGET SEQUENCES FROM DEGRADATION - Methods are provided for selective tagging of short nucleic acids comprising a short target nucleotide sequence over longer nucleic acids comprising the same target nucleotide sequence. The methods can involve performing one or two cycles of amplification of a sample comprising long nucleic acids and short nucleic acids, each comprising the same target nucleotide sequence with at least two target-specific primers or primer pairs under suitable annealing conditions, wherein the primer pairs comprise: an inner primer or primer pair that can amplify the target nucleotide sequence on long and short nucleic acids (wherein each inner primer comprises a 5′ nucleotide tag; and an outer primer or primer pair that amplifies the target nucleotide sequence on long nucleic acids, but not on short nucleic acids); whereby the amplification after a second cycle produces at least one tagged target nucleotide sequence that comprises two nucleotide tags, one from each inner primer, with the target nucleotide sequence located between the nucleotide tags.11-11-2010
20100285538METHODS FOR NUCLEIC ACID MANIPULATION - A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.11-11-2010
20100285536METHOD AND APPARATUS FOR AMPLIFICATION OF NUCLEIC ACID SEQUENCES BY USING THERMAL CONVECTION - The present invention provides a nucleic acid sequence amplification method and apparatuses thereof that are simple in the design and easy to miniaturize and integrate into complex apparatuses, with capability of using DNA polymerases that are not thermostable. In the present invention, a plurality of heat sources are combined to supply or remove heat from specific regions of the sample such that a specific spatial temperature distribution is maintained inside the sample by locating a relatively high temperature region lower in height than a relatively low temperature region.11-11-2010
20100285535PCR HOT START BY MAGNESIUM SEQUESTRATION - The present invention is directed to a synthetic peptide having a length of not more than 30 amino acids comprising a divalent cation binding site. Such a peptide according to the present invention is part of a composition for nucleic acid amplification and provides for a so-called hot start effect.11-11-2010
20100015667METHOD OF IN VITRO POLYNUCLEOTIDE SEQUENCES SHUFFLING BY RECURSIVE CIRCULAR DNA MOLECULES FRAGMENTATION AND LIGATION - The present invention relates to a method for in vitro recombining polynucleotides sequences of interest from at least two initial different polynucleotide sequences of interest by cleavage/ligation of circular DNA molecules.01-21-2010
20110136180LYSIS AND REVERSE TRANSCRIPTION FOR MRNA QUANTIFICATION - The present invention is directed to a method for performing an RT-PCR for amplifying a target RNA comprising the steps of a) lysis of a cellular sample which is supposed to contain the target RNA with a lysis buffer comprising between 0.2 M and 1 M Guanidine Thiocyanate, b) diluting the sample to an extend such that Guanidine Thiocyanate is present in a concentration of about 30 to 50 mM, c) reverse transcribing in the presence of a mixture of first strand cDNA synthesis primers, the mixture consisting of oligo dT primers and random primers, and d) subjecting the sample to multiple cycles of a thermocycling protocol and monitoring amplification of the first strand cDNA in real time, characterized in that steps a) to c) are done in the same vessel.06-09-2011
20100304443SAMPLING DEVICE - The present invention generally relates to devices, systems, and methods for acquiring and/or dispensing a sample without introducing a gas into a microfluidic system, such as a liquid bridge system. An exemplary embodiment provides a sampling device including an outer sheath; a plurality of tubes within the sheath, in which at least one of the tubes acquires a sample, and at least one of the tubes expels a fluid that is immiscible with the sample, in which the at least one tube that acquires the sample is extendable beyond a distal end of the sheath and retractable to within the sheath; and a valve connected to a distal portion of the sheath, in which the valve opens when the tube extends beyond the distal end and closes when the tube retracts to within the sheath.12-02-2010
20100323405Droplet-Based Nucleic Acid Amplification in a Temperature Gradient - A method of amplifying a nucleic acid, the method comprising cycling an amplification-ready droplet through a temperature gradient to locations within the temperature gradient suitable for effecting steps in an amplification reaction.12-23-2010
20100323404METHOD FOR RECOMBINING DNA SEQUENCES AND COMPOSITIONS RELATED THERETO - Provided herein are methods for manipulating nucleotide sequences that permit greater control of sequence recombination compared to traditional methods, and related compositions. In one such method, a set of oligonucleotides and at least one primer are provided, where the primer has a first region uniquely complementary to a sequence of a first oligonucleotide of the set and a second region uniquely complementary to a second oligonucleotide of the set, combining the primer with an oligonucleotide comprising the first region of said first polynucleotide and an oligonucleotide comprising the second region of said second polynucleotide, and PCR amplifying to create a chimeric polynucleotide having some sequence from the first oligonucleotide and some sequence from the second oligonucleotide.12-23-2010
20110076726Differential enzymatic fragmentation by whole genome amplification - The present invention provides methods for detecting the presence of methylation at a locus within a population of nucleic acids.03-31-2011
20090176281MODULAR VECTOR SYSTEMS - The present invention provides improved techniques and reagents for producing nucleic acid molecules. In certain preferred embodiments, the nucleic acid molecules are modular vectors. In certain preferred embodiments, the nucleic acid molecules are produced in polymerase chain reactions employing terminator primer residues.07-09-2009
20090197306THERMAL CYCLING DEVICE - A multi-layer device suitable for thermal cycling is set forth. The device is particularly suitable for performing a polymerase chain reaction (PCR). One embodiment includes a first layer (08-06-2009
20090176282Device and Method for Thermal Cycling - A thermal cycling device for performing nucleic acid amplification on a plurality of biological samples positioned in a sample well tray. The thermal cycling device includes a sample block assembly, an optical detection system, and a sample well tray holder configured to hold the sample well tray. The sample block assembly is adapted for movement between a first position permitting the translation of the sample well tray into alignment with sample block assembly, and a second position, upward relative to the first position, where the sample block assembly contacts the sample well tray. A method of performing nucleic acid amplification on a plurality of biological samples positioned in a sample well tray in a thermal cycling device is also provided.07-09-2009
20110262973Methods and Compositions for DNA Manipulation - single-stranded extension having a desired length and sequence composition. Methods for forming single-stranded extensions include: the use of a cassette containing at least one nicking site and at least one restriction site at a predetermined distance from each other and in a predetermined orientation; or primer-dependent amplification which introduces into a polynucleotide molecule, a modified nucleotide which is excised to create a nick using a nicking agent. The methods and compositions provided can be used to manipulate a DNA sequence including introducing site specific mutations into a polynucleotide molecule and for cloning any polynucleotide molecule or set of joined polynucleotide molecules in a recipient molecule such as a vector of choice.10-27-2011
20100190215PENETRABLE CAP - A cap having a core structure dimensioned to receive a pipette therethrough. The cap includes two axially aligned frangible seals that are affixed to the core structure in a spaced-apart relationship. The frangible seals include a foil layer and are constructed so that air passageways are formed between the frangible seals and a pipette tip when the pipette tip penetrates the frangible seals. The cap optionally includes a filter interposed between the first and second frangible seals.07-29-2010
20100190214BIOCHEMICAL TREATMENT APPARATUS AND METHOD COMPRISING LIQUID HANDLING MECHANISM - Nucleic acid solution, magnetic particles solution and primer or eluate are moved between a lot of wells provided in reaction/storage vessel 07-29-2010
20100028955Sequence Amplification with Target Primers - The present disclosure relates to the amplification of target nucleic acid sequences for various sequencing and/or identification techniques. This can be accomplished via the use of target primers and isothermal multiple strand displacement (MDA) processes. The use of these target primers and MDA, as described herein, allows for, for example, the reduction in the amplification of undesired hybridization events (such as primer dimerization and the “jackpot mutation” effect of PCR) while allowing for the amplification of the target nucleic acid sequences.02-04-2010
20100021972METHOD AND APPARATUS FOR AMPLIFYING NUCLEIC ACIDS - A method and apparatus for amplifying nucleic acids. The method includes introducing into a reaction vessel via different inlet channels a reactant aqueous solution containing reactants for nucleic acid amplification and a fluid that is phase-separated from the reactant aqueous solution and does not participate in amplification reaction, creating a plurality of reactant aqueous solution droplets surrounded by the fluid by contacting the reactant aqueous solution with the fluid in the reaction vessel, and amplifying the nucleic acids in the reactant aqueous solution droplets. The apparatus includes a substrate, a reaction vessel formed inside of the substrate, at least one first inlet channel formed inside the substrate, connected to an end of the reaction vessel, and allowing introduction of a reactant aqueous solution containing reactants for nucleic acid amplification into the reaction vessel, a second inlet channel formed inside the substrate, connected to the end of the reaction vessel, and allowing introduction of a fluid that is phase-separated from the reactant aqueous solution and does not participate in amplification reaction into the reaction vessel, and a heating unit installed on the substrate in such a way to thermally contact with the substrate and heating the substrate.01-28-2010
20100021970Nucleic Acid Amplification Using a Reversibly Modified Oligonucleotide - The present invention provides a method for amplification of a target nucleic acid sequence or signal, wherein an amplification reaction mixture is used which contains at least one reversibly modified oligonucleotide having a non-hydroxyl group 3′ end which can be converted into a hydroxyl 3′ end upon exposure to a chemical and/or irradiation and/or a range of temperature. The present invention also provides a reversibly modified oligonucleotide as described above, and a nucleic acid amplification reaction mixture and kit comprising such an oligonucleotide.01-28-2010
20100021971METHOD TO REMOVE REPETITIVE SEQUENCES FROM HUMAN DNA - The invention discloses an innovative method to deplete repetitive sequences from human DNA. The method comprises (a) providing a source DNA containing both unique and repetitive sequences and sonicating the source DNA to smaller fragments; (b) providing a driver DNA containing sequences complementary to the repetitive sequences of the source DNA and labeled with a non-radioactive label, (c) hybridizing the source DNA and the driver DNA in the presence of a molecule that binds the label to form a complex; (d) removing the hybridized repetitive sequences from the complex by using RNAase and electrophoresis or by incubating with a mixture of phenol, chloroform, and ethanol; and (e) recovering the remaining source DNA wherein said repetitive sequences being significantly removed.01-28-2010
20090263870SYSTEM AND METHOD FOR AMPLIFYING A NUCLEIC ACID MOLECULE - There is provided a method and/or system which allow on-chip preconditioning of complex real-world samples and/or handling of limited amounts of target material, and/or on-chip nucleic acid amplification process, using a free droplet containing magnetic attractable material. The nucleic acid amplification process comprises controlling the position of the magnetic attractable material and performing the nucleic acid amplification in a thermocycling droplet located onto at least one temperature zone. The low thermal masses of the herein described heaters/temperature sensors come along with fast temperature transitions within the corresponding temperature zones allowing impressing temperature gradients in at least one temperature zone between subsequent or within the same thermocycle(s). Additionally, the variable residence times of the droplet in a given temperature zone permit to customize the denaturation, annealing and/or extension times within the same or between different PCR runs. Additionally, the herein described method and/or system allow amplification of one or more nucleic acid molecules. Additionally, the herein described method and/or system allow real-time monitoring with or without the presence of magnetic attractable material bound to said nucleic acid molecule.10-22-2009
20090263871Methods and Compositions for Amplification of DNA - The invention provides an Enzyme Blend comprising a DNA polymerase and a DNA repair enzyme. Methods and kits for amplification of DNA that is damaged, undamaged, or suspected of being damaged are also provided.10-22-2009
20100240102METHODS AND COMPOSITIONS FOR MULTIPLE DISPLACEMENT AMPLIFICATION OF NUCLEIC ACIDS - Disclosed are methods for multiple displacement amplification of a nucleic acid sequence in a sample. The nucleic acid is contacted with a reaction mixture that includes a set of oligonucleotide primers and a plurality of polymerase enzymes. The reaction mixture is subjected to conditions under which the nucleic acid sequence is amplified to produce an amplification product in a multiple displacement amplification reaction. Also disclosed are kits containing a set of oligonucleotide primers with random sequences having lengths of 6 to 8 nucleobases. At least some of the individual members of the primers have one or more ribose modifications that stabilize or lock the ribose ring in a 3′ -endo conformation. At least some of the primers have one or more universal nucleobases.09-23-2010
20090023190SEQUENCE AMPLIFICATION WITH LOOPABLE PRIMERS - The present disclosure relates to the amplification of target nucleic acid sequences. This can be accomplished via the use of various primers. The use of these primers, as described herein, results in nucleic acid structures that can reduce the amplification of nonspecific hybridization events (such as primer dimerization) while allowing the amplification of the target nucleic acid sequences.01-22-2009
20080274512Reagent Transfer Device - A device for transferring reagents in solid or liquid form from one place to another on an apparatus, said device comprising an upper portion, adapted to be held, and a lower portion adapted to hold reagents in solid or liquid form, in an available manner. For example, the reagents may be held within a cage structure or retained on an outer surface of the lower portion. Methods for using the device, as well as apparatus for use in conjunction with the device are also described and claimed.11-06-2008
20120231509Primers and Probes for the Amplification and Detection of HIV GAG, REV and NEF Polynucleotides - The invention relates to improved methods and compositions for the nucleic acid amplification of one or multiple variants (strains) of Human Immunodeficiency Virus (HIV) present in a sample, and preferably in a sample from a pathogen infected individual. In particular, novel primers, methods and kits for the amplification of one or more species of HIV Rev, Gag and Nef nucleic acids are provided. The amplified HIV nucleic acid can be used to identify and/or quantitate HIV variants present in a sample. Nucleic acids produced by the methods of the invention or the proteins encoded thereby can also be used directly as vaccines or to transfect/load antigen presenting cells. The loaded antigen presenting cells can be used as a vaccine for the treatment or prevention of HIV infection.09-13-2012
20100203594High-speed pcr using high-speed dna polymerase - [Problems] To achieve a high-speed PCR which can be completed in a period of several minutes and thus can be used for a new application.08-12-2010
20110008845PRIMERS FOR PCR AMPLIFICATION COMPRISING A BASIC PARTS WITHIN THE PRIMER SEQUENCES - The present invention relates to primers for PCR amplification comprising abasic parts within the primer sequences and a method for PCR amplification using the same. More precisely, the present invention relates to primers capable of amplifying different templates and having abasic parts complementary to mutated site or polymorphic site of template DNA and a method for PCR amplification comprising the steps of mixing the composition for PCR amplification comprising the primers with nucleic acid template; and performing PCR with the mixture. The primers for PCR amplification of the present invention contain abasic parts not having specific coding information in their nucleotide sequences, so that they can amplify different templates having mutated sites at the same time.01-13-2011
20090035825Devices, Systems, and Methods for Preparing Emulsions - A vortex mixer and method for forming an emulsion wherein the mixer is adapted to form an emulsion with a desired droplet size and having a desired volume. The vortex mixer provides improved uniformity in emulsion preparation and may be used to create multiple emulsions simultaneously.02-05-2009
20090176280Amplification and cloning of single dna molecules using rolling circle amplification - The present invention relates, e.g., to a method for amplifying a small number of copies (e.g. a single copy) of a single-stranded circular DNA molecule (e.g. having a size of about 5-6 kb) by an isothermal rolling circle mechanism, using random or partially random primers and a F29-type DNA polymerase. The method, which can also be used for amplifying DNAs by non-rolling types of multiple displacement amplification, comprises incubating the reaction components in a small volume, e.g. about 10 μl or less, such as about 0.6 μl or less. The degree of amplification can be about 109 fold, or higher. A method for cell-free cloning of DNA, using the rolling circle amplification method of the invention, is described.07-09-2009
20080220481Sample Plate Assembly and Method of Processing Biological Samples - The invention concerns a v-bottomed sample plate, a frame for sample plates and a kit and method for processing biological samples. The kit comprises a tray assembly and a plurality of sample plates designed to fit into the tray assembly. The tray assembly comprises a frame having a central plate receiving portion having a width and length, whereby said tray assembly is capable of accommodating the sample plates side by side in the plate receiving portion. Each of the sample plates contains a plurality of individual sample wells arranged in a grid, the dimension of the plate in a first direction being at maximum the width of the frame and the dimension of the plate in a second direction being at maximum half of the length of the plate receiving portion of the of the frame, and means for enabling automated handling of the plates. The invention enables more efficient biomedical processing of samples.09-11-2008
20120301924METHOD FOR INCUBATING THE CONTENTS OF A RECEPTACLE - In a computer-controlled, automated method, a nucleic acid amplification procedure is performed within a prescribed incubation temperature range within an instrument that comprises at least one thermal element situated at a first location of the instrument and an amplification incubator situated at a second, spaced-apart location of the instrument. A receptacle and its contents are exposed to the thermal element for a predetermined period of time at the first location, such that the temperature of the contents of the receptacle is adjusted to approximate the incubation temperature. The receptacle is then moved from the first location and into the amplification incubator, and, in the amplification incubator, a nucleic acid contained in the contents of the receptacle is subjected to the nucleic acid amplification procedure.11-29-2012
20100173365DNA polymerase fusions and uses thereof - The present invention discloses methods of using DNA polymerase fusions at high pH in PCR, DNA sequencing and mutagenesis protocols.07-08-2010
20120040404THERMAL CYCLING DEVICE - Multi-layer devices suitable for thermal cycling processes. The devices are particularly suitable for performing polymerase chain reactions (PCR). One embodiment includes a first conducting layer, a second conducting layer adjacent to the first layer, and a third conducting layer adjacent to the second layer opposite the first layer. Insulating layers are positioned between said three conducting layers. Continuous channels are formed within the layers. The channels can be formed in either the conducting layer or the insulating layers, or both. Other embodiments include two conducting layers. At least one integral or separate temperature source may be provided to maintain the conducting layers at various desired temperatures.02-16-2012
20110318785DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.12-29-2011
20110318786DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.12-29-2011
20110318784PREPARATION AND AMPLIFICATION OF NUCLEIC ACIDS BY MEANS OF MAGNETIC PARTICLES - The invention relates to the preparation of a biological sample for performing verifications and examinations, wherein the aim of the invention is the creation of a method for preparing a biological sample having an improved PCR sensitivity compared to the reference standard having standard PCR without having to raise the cost thereof.12-29-2011
20120045798METHOD AND DEVICE FOR ELECTROWETTING GENETIC ANALYSIS - It relates to a method for amplifying at least one nucleotide molecule contained in a sample (E) comprising the following successive steps consisting of (a) subjecting said sample (E) to a 1st amplification step on an area (Z02-23-2012
20120045796NUCLEIC ACID HOTSTART TECHNOLOGY - The present invention relates to molecules used as hotstarts for polymerases, including among others the DNA polymerase and reverse transcriptase.02-23-2012
20120003699SEPARTION OF TAGGED FRAGMENTS - Methods of separating tagged nucleic acid fragments from populations of fragments are provided. The fragments are produced from a population of tagged nucleic acids. In some cases, the population of tagged nucleic acids comprises a population of clonal nucleic acids.01-05-2012
20120045797NUCLEIC ACID AMPLIFICATION PROCEDURE - The invention provides methods for amplification of polynucleotide sequences using primers containing single-Cstranded RNA. The methods employ use of an enzyme capable of cleaving single-stranded RNA, such as RNase I, to degrade a first RNA-containing primer prior to addition of a second RNA-containing primer. The invention also provides compositions and kits pl. for practicing the amplification methods, as well as methods which use the amplification products.02-23-2012
20120003698LINEAR AMPLIFICATION OF SHORT NUCLEIC ACIDS - The present teachings provide novel methods for amplifying short nucleic acids. In some embodiments, the present teachings provide novel methods for linearly amplifying a collection of micro RNAs by using temperature cycling during a reverse transcription reaction. The cycling can comprise at least 20 cycles of an annealing temperature segment of 10° C.-30° C., and a denaturation temperature segment of 35° C.-60° C. In some embodiments, the temperature cycled reaction can comprise an osmolyte.01-05-2012
20120208240DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.08-16-2012
20120208241THERMOCYCLING DEVICE FOR NUCLEIC ACID AMPLIFICATION AND METHODS OF USE - The present invention provides thermocycling devices useful for amplification of nucleic acids in droplets. The thermocycling device utilizes the flow of one or more fluids through a main compartment at temperatures sufficient to conduct a polymerase chain reaction. Methods of amplifying nucleic acids in droplets are also provided.08-16-2012
20120208239DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.08-16-2012
20120058517Recombinase Polymerase Amplification - This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further disclosed are conditions to enable real-time monitoring of RPA reactions, methods to regulate RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carry-over contamination, and methods to employ sequence-specific third ‘specificity’ probes. Further described are novel properties and approaches for use of probes monitored by light in dynamic recombination environments.03-08-2012
20120009629METHODS AND KITS FOR USE IN THE SELECTIVE AMPLIFICATION OF TARGET SEQUENCES - The present invention provides nucleic acid amplification methods that desirably reduce or eliminate false positive amplification signals resulting from contaminating biological material, e.g., nucleic acid, that may be present in one or more reagents used in an amplification reaction and/or that may be present in the environment in which an amplification reaction is performed. The invention offers the further advantage of requiring less stringent purification and/or sterility efforts than conventionally needed in order to ensure that enzymes and other reagents used in amplification reactions, and the environment in which an amplification reaction is performed, are free of bacterial or other nucleic acid contamination that may yield false positive results.01-12-2012
20120009628DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.01-12-2012
20100129875EMULSION PCR AND AMPLICON CAPTURE - Compositions and methods of use are disclosed for clonally amplifying target polynucleotide sequences in solution and attaching the amplicons to a surface by activation of a masked binding moiety.05-27-2010
20090117622MIXTURE OF REVERSIBLY INHIBITED ENZYMES - The invention relates to composition or a kit containing an enzyme that is reversibly inhibited by means of a chemical modification and an enzyme which is reversibly inhibited using non-covalent binding, the use of a mixture of enzymes reversibly inhibited in such a manner for processing or multiplying polynucleotides, and a method for specifically amplifying DNA by simultaneously using both types of reversibly inhibited enzymes.05-07-2009
20110165628THERMOCYCLING DEVICE - The present invention is related to a thermocycling device (07-07-2011
20110165627GENE SITE SATURATION MUTAGENESIS - A method for producing progeny polynucleotides and polypeptides by Gene Site Saturation Mutagenesis (GSSM). The method provides a set of degenerate primers corresponding to codons of a template polynucleotide, and performs polymerase elongation to produce progeny polynucleotides, which contain sequences corresponding to the degenerate primers. The progeny polynucleotides can be expressed and screened for directed evolution.07-07-2011
20120058516SEALING MULTIWELL PLATES - The present invention relates to a heat sealable member comprising a sheet of heat sealable material to a multiwell plate, wherein the sheet is mounted in a frame, and wherein said frame is sized to fit over a multiwell plate so that the sheet is contacted with at least one of the rims of the top of the plate and the individual wells; as well as to the use of the heat sealable member with automatic plate handling apparatus, especially in PCR processes. The present invention further relates to a method for sealing multiwell plates.03-08-2012
20120058518NUCLEIC ACID ISOTHERMAL AMPLIFICATION METHOD - A nucleic acid isothermal amplification method includes: performing a reverse transcription reaction to reverse-transcribe a target RNA strand into a template DNA strand; irradiating a reaction solution with light to dissociate a photodegradable protecting group bound to a nucleotide in a sequence of an oligonucleotide primer; and performing an amplification reaction for the template DNA strand.03-08-2012
20120058515METHOD OF REGULATING OLIGONUCLEOTIDE FUNCTIONALITY - The present invention relates generally to a method of regulating oligonucleotide functionality and, more particularly, to a method of regulating the functionality of a primer or probe. The method of the invention is designed to provide a means to selectively inactivate or activate the functionality of an oligonucleotide, such as a primer, thereby providing means to regulate the progress of any method using that oligonucleotide. The development of a means to regulate the functionality of an oligonucleotide, such as a primer, is useful in a range of applications including, but not limited to, amplification reactions such as PCR, isothermal amplification and nucleic acid strand extension. With respect to amplification reactions, these have wide utility including the diagnosis and/or monitoring of disease conditions which are characterised by specific gene sequences and the characterisation or analysis of specific gene regions of interest.03-08-2012
20120058519METHOD, DEVICES, AND SYSTEMS FOR FLUID MIXING AND CHIP INTERFACE - In one aspect, the present invention provides methods, devices, and systems for ensuring that multiple components of a mixture are fully mixed in a continuous flow microfluidic system while ensuring that mixing between segments flowing through the chip is minimized. In some embodiments, the present invention includes mixing fluids in a droplet maintained at the tip of a pipette before the mixture is introduced to the microfluidic device. In another aspect, the present invention provides methods, devices, and systems for creating segments that move through a microfluidic chip with minimal mixing between segments. The microfluidic chip may have an interface chip and a reaction chip. In some embodiments, the present invention includes creating segments that flow through an interface chip and a reaction chip, wherein the interface chip and a reaction chip have separate flow control mechanisms and produce minimal mixing between segments.03-08-2012
20120064576METHOD AND COMPOSITION FOR ENHANCING EFFICIENCY AND SENSITIVITY IN POLYMERASE CHAIN REACTION - Methods and compositions for enhancing reaction efficiency and sensitivity in polymerase chain reaction (PCR) are disclosed.03-15-2012
20120156729SELECTIVE TERMINAL TAGGING OF NUCLEIC ACIDS - Methods are provided for adding a terminal sequence tag to nucleic acid molecules for use in RNA or DNA amplification. The tag introduced may be used as a primer binding site for subsequent amplification of the DNA molecule and/or sequencing of the DNA molecule and therefore provides means for identification and cloning of the 5′-end or the complete sequence of mRNAs.06-21-2012
20100120098TRANSPOSON END COMPOSITIONS AND METHODS FOR MODIFYING NUCLEIC ACIDS - The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5′-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5′- and 3′-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5′-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3′-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5′- and 3′-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called “next-generation sequencing.)05-13-2010
20110086392Methods for Repairing Degraded DNA - The present invention provides methods and kits for repair of degraded DNA which may then be used as a template for efficient amplification by a number of different amplification reactions. The method relies upon a series of enzymatic activities provided by DNA repair enzymes.04-14-2011
20130011880AUTOMATED ENRICHMENT FOR NUCLEIC ACID SEQUENCING - The present teachings provide apparatuses and methods for automated handling of samples, e.g., biological or chemical samples. The apparatuses and the methods of the present teachings allow automated performance of various sample manipulation steps without manual intervention. In a preferred embodiment, the present teachings provide apparatuses and methods for automated enrichment of templated beads produced by PCR.01-10-2013
20110065151NUCLEIC ACID AMPLIFICATION WITH SINGLE STRAND DNA BINDING PROTEIN - A method is disclosed in which circular DNA molecules are amplified preferentially in a mixture of circular DNA molecules and linear DNA molecules by the inclusion of single strand DNA binding protein.03-17-2011
20110065150REACTION VESSEL COMPRISING ELECTRICALLY CONDUCTING POLYMER AS A HEATING ELEMENT - A reaction vessel for conducting a chemical or biochemical reaction, such as a polymerase chain reaction wherein electrically conducting polymer is arranged to act as a heating element. The profile of the electrically conductive polymer differs in different regions of the vessel so as to control thermal gradients. The profile of the electrically conductive polymer may be arranged to either increase or reduce the thermal gradient. Reaction systems comprising combinations of vessels of the invention and apparatus for heating them, as well as particular reactions vessels are also described and claimed.03-17-2011
20110104760METHODS AND COMPOSITIONS FOR IMPROVING EFFICIENCY OF NUCLEIC ACIDS AMPLIFICATION REACTIONS - The present invention provides methods and compositions for improving the efficiency of nucleic acid amplification reactions. The invention encompasses hybrid polymerases that show increased processivity over wild type polymerases as well as decreased exonuclease activity. The invention also encompasses methods, compositions and kits for conducting nucleic acid synthesis and amplification reactions in which non-specific amplification of primers is reduced.05-05-2011
20110104763Sequential Addition of Short DNA Oligos In DNA-Polymerase-Based Synthesis Reactions - A method of fabricating a DNA molecule of user-defined sequence. The method comprises the steps of preselecting a multiplicity of DNA sequence segments that will comprise the DNA molecule of user-defined sequence, separating the DNA sequence segments temporally, and combining the multiplicity of DNA sequence segments with at least one polymerase enzyme wherein the multiplicity of DNA sequence segments join to produce the DNA molecule of user-defined sequence. Sequence segments may be of length n, where n is an even or odd integer. In one embodiment the length of desired hybridizing overlap is specified by the user and the sequences and the protocol for combining them are guided by computational (bioinformatics) predictions. In one embodiment sequence segments are combined from multiple reading frames to span the same region of a sequence, so that multiple desired hybridizations may occur with different overlap lengths. In one embodiment starting sequence fragments are of different lengths, n, n+1, n+2, etc.05-05-2011
20110104762Detection probe acting by molecular recognition - The present invention relates to a detection probe acting by molecular recognition of a target sequence, comprising successively in the 5′-3′ direction: 05-05-2011
20110104761THERMOSTABLE DNA POLYMERASE FROM PALAEOCOCCUS HELGESONII - There is provided a polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 78% identity to 05-05-2011
20090162903NUCLEIC ACID AMPLIFICATION METHOD - An object to be achieved by the present invention is to provide a nucleic acid amplification method by which a nucleic acid can be amplified using oligonucleotide primers and DNA polymerase. The present invention provides a nucleic acid amplification method which comprises performing incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3′ end of the primer and thus amplifying the nucleic acid fragment, wherein a tag sequence is added at the 5′ end of the first oligonucleotide primer, and the tag sequence is a nucleotide sequence on the template nucleic acid fragment which is present downstream of the sequence which is substantially complementary with the 3′ end region of the first oligonucleotide primer (a region where the first oligonucleotide is annealed to the template nucleic acid).06-25-2009
20120122160THERMAL CYCLER AND THERMAL CYCLING METHOD - A thermal cycler includes a holder to which a biotip having a longitudinal direction is attached in such a manner that one end portion of the biotip is at a higher level than the other end portion, and that the distance between one end portion of the biotip and the rotational axis is shorter than the distance between the other end portion of the biotip and the rotational axis, a heating unit heats a first end portion of the biotip, a rotating unit rotates the holder, and a controller that controls the rotation speed of the rotating unit. The controller has a first mode a rotation speed at which the magnitude of the centrifugal force acting on the reaction mixture becomes smaller than the gravity, and a second mode a rotation speed at which the magnitude of the centrifugal force acting on the reaction mixture becomes greater than the gravity.05-17-2012
20120122159PCR-BASED METHOD OF SYNTHESIZING A NUCLEIC ACID MOLECULE - There is provided a method of synthesizing a nucleic acid molecule and in one aspect, the method comprises assembling a full length template nucleic acid molecule by PCR in a PCR reaction mixture comprising a set of assembly oligonucleotides having a first average melting temperature and a set of outer amplification primers having a second average melting temperature that is lower than the first average melting temperature, wherein said assembling comprises subjecting the PCR reaction mixture to a first annealing temperature that is higher than the second average melting temperature and; amplifying the full length template nucleic acid molecule by PCR in the PCR reaction mixture wherein said amplifying comprises subjecting the PCR reaction mixture to a second annealing temperature that permits annealing of the outer amplification primers to the full length template nucleic acid molecule.05-17-2012
20120122158PCR FOR DNA THAT IS RESISTANT TO AMPLIFICATION - DNA that is difficult to amplify by conventional PCR is amplified by a modified PCR process that includes a high-temperature, short-term heating step as the last step of each thermal cycle, following the conventional denaturing and annealing/elongation steps.05-17-2012
20090130718Gene site saturation mutagenesis - A method for producing progeny polynucleotides and polypeptides by Gene Site Saturation Mutagenesis (GSSM). The method provides a set of degenerate primers corresponding to codons of a template polynucleotide, and performs polymerase elongation to produce progeny polynucleotides, which contain sequences corresponding to the degenerate primers. The progeny polynucleotides can be expressed and screened for directed evolution.05-21-2009
20100248310METHOD OF DNA AMPLIFICATION - The present invention relates generally to a method of amplifying a nucleic acid region of interest and, more particularly, to a method of amplifying a nucleic acid region of interest using a PCR method designed to minimise the generation of amplicons from primers which have bound to nucleic acid regions other than the specific region of interest. The method of the present invention is based on the determination that by rendering inefficient the functionality of either the forward primer or the reverse primer, the rate of amplification of irrelevant nucleic acid regions can be reduced relative to amplification of the region of interest. The provision of a selective means of amplifying a nucleic acid region of interest is useful in a range of applications including, but not limited to, the diagnosis and/or monitoring of disease conditions which are characterised by specific gene sequences, the characterisation or analysis of gene regions of interest, the identification or characterisation of DNA breakpoint regions and the isolation of gene sequences of interest where only the nucleotide sequence at one end of the gene sequence of interest is known.09-30-2010
20110183378NUCLEIC ACID AMPLIFICATION METHOD, NUCLEIC ACID AMPLIFICATION APPARATUS, AND CHIP USED IN NUCLEIC ACID AMPLIFICATION - A nucleic acid amplification method includes introducing a liquid sample into a first chamber of a chip for use in nucleic acid amplification, the chip including a second chamber containing liquid that has a smaller specific gravity than the liquid sample and is immiscible with the liquid sample, injecting the liquid sample into the second chamber from the first chamber by a centrifugal force, regulating a temperature of an end of the chip, and rotating the chip about a rotation axis at a predetermined speed.07-28-2011
20120164692ANNEALING CONTROL PRIMER AND ITS USES - The present invention relates to an annealing control primer for improving annealing specificity in nucleic acid amplification and its applications to all fields of nucleic acid amplification-involved technology. The present primer comprises (a) a 3′-end portion having a hybridizing nucleotide sequence substantially complementary to a site on a template nucleic acid to hybridize therewith; (b) a 5′-end portion having a pre-selected arbitrary nucleotide sequence; and (c) a regulator portion positioned between said 3′-end portion and said 5′-end portion comprising at least one universal base or non-discriminatory base analog, whereby said regulator portion is capable of regulating an annealing portion of said primer in association with annealing temperature.06-28-2012
20120164690UNIVERSAL REFERENCE DYE FOR QUANTITATIVE AMPLIFICATION - Universal reference dye mixtures for quantitative amplification, and uses thereof, are provided.06-28-2012
20100209971GENERATION OF NUCLEIC ACID MOLECULES - The present invention relates generally to methods for generating single stranded nucleic acid molecules following enhanced solid phase polynucleotide amplification. The present invention employs an amplification reaction using primers with differential priming properties at particular annealing conditions or an immobilized primer nested between two aqueous phase primers. Thus, by primer design, solid support primer participation is enhanced relative to aqueous phase primers. The subject invention further provides methods for labeling solid matrices with single and double stranded nucleic acid molecules. Kits for generating single stranded nucleic acid molecules and for conducting amplification reactions also form part of the present invention. The present invention further provides amplification systems for the generation of single stranded nucleic acid molecules optionally labelled with a reporter molecule and their use inter alia as labels, primers and probes.08-19-2010
20100209972METHOD FOR SYNTHESIS OF SINGLE- OR DOUBLE-STRANDED DNA, AND KIT FOR THE SYNTHESIS - An object of the present invention is to provide a simple and safe method for synthesis of single-stranded or double-stranded DNA and a kit for performing the synthesis. The present invention relates to, (1) a method for synthesis of single-stranded DNA, comprising a nucleotide sequence corresponding to the template RNA, characterized by comprising the following steps: 1) Step 1 in which the template RNA is subjected to reverse transcription reaction, 2) Step 2 in which the solution obtained in the treatment of Step 1 is subjected to alkaline treatment; (2) a method for synthesis of double-stranded DNA, comprising a nucleotide sequence corresponding to the template RNA, characterized by comprising the following steps: 1) Step 1 in which the template RNA is subjected to reverse transcription reaction, 2) Step 2 in which the solution obtained in the treatment of Step 1 is subjected to alkaline treatment, 3) Step 3 in which the obtained single-stranded DNA is converted to a double strand, as well as (3) a kit for synthesis of single-stranded DNA and (4) a kit for synthesis of double-stranded DNA for use in the above-described method for synthesis.08-19-2010
20120214207METHODS AND SYSTEMS FOR FAST PCR HEATING - A microplate for polymerase chain reaction (PCR) comprises a substrate having a metallic material for heating PCR samples, and a barrier layer disposed adjacent to the substrate. In some cases, the barrier layer is formed of a first polymeric material. The microplate includes one or more wells for containing PCR samples. The one or more wells are formed of a second polymeric material sealed to the barrier layer. In some cases, the substrate can provide a PCR ramp rate of at least about 5° C./second.08-23-2012
20120135471Method for Concentrating Sample Constituents and Amplifying Nucleic Acids - A method is disclosed for concentrating sample constituents and for multiplying nucleic acids from a biological sample which are containing in the sample constituents. The nucleic acids are amplified on the same filter on which the sample constituents are also separated off.05-31-2012
20120135473PROCESSES USING DUAL SPECIFICITY OLIGONUCLEOTIDE AND DUAL SPECIFICITY OLIGONUCLEOTIDE - The present invention relates to various processes by a template-dependent extension reaction using a dual specificity oligonucleotide and a dual specificity oligonucleotide composed of three different Tm portions therefor. Demonstrated in the present invention are the features of the dual specificity oligonucleotide, which are high hybridization specificity and mismatch tolerance.05-31-2012
20120135472HOT-START PCR BASED ON THE PROTEIN TRANS-SPLICING OF NANOARCHAEUM EQUITANS DNA POLYMERASE - Disclosed is a hot-start PCR method, based on protein trans-splicing of intein-inserted large (Neq L) and small (Neq S) fragments of Neq DNA polymerase. The method comprises: preparing a PCR reaction mixture containing a sample DNA and primers; adding the Neq L fragment and the Neq S fragment together to the PCR reaction mixture, said Neq L fragment consisting of an amino acid sequence of SEQ ID NO: 2, with an intein amino acid sequence stretching from position 579 to 676 therein, said Neq S fragment consisting of an amino acid sequence of SEQ ID NO: 4 with an intein amino acid sequence stretching from position 1 to 30 therein; inducing the Neq L fragment and the Neq S fragment to undergo a protein trans-splicing process to form a polypeptide exhibiting Neq DNA polymerase activity; and performing a certain number of cycles of DNA denaturation, primer annealing and DNA extension.05-31-2012
20100173364Repair of Nucleic Acids for Improved Amplification - Methods and compositions are provided for repairing a polynucleotide so that it can be copied with improved fidelity and/o yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a DNA ligase and an effective amount of at least one endonuclease as well as a cofactor selected from NAD07-08-2010
20100173363Use of Single-Stranded Nucleic Acid Binding Proteins in Sequencing - The invention provides methods for stabilizing a nucleic acid sequencing reaction. Generally, methods of the invention include exposing a target nucleic acid to a single-stranded nucleic acid binding protein and performing a sequencing reaction.07-08-2010
20120171726RAPID WHOLE GENOME AMPLIFICATION - The present invention provides compositions and methods for rapidly amplifying target nucleic acid (e.g., using whole genome amplification) that allows small amounts of starting nucleic acid to be employed. In certain embodiments, the methods employ compositions that comprise: phi29 polymerase, exo- Klenow polymerase and/or Klenow polymerase, dNTPs, primers, and a buffering agent. In some embodiments, the target nucleic acid is amplified at a rate that would result in at least 1000-fold amplification in thirty minutes.07-05-2012
20120171728PROCESS FOR AMPLIFYING DNA USING TETRATETHYLENE GLYCOL, KIT OF PARTS THEREFOR AND USE THEREOF - Disclosed is a process for amplifying DNA, a process for preparing and amplifying DNA, a kit of parts for DNA amplification and DNA preparation, and the use thereof, all of which are characterized by the use of tetraethylene glycol. Moreover, the use of tetraethylene glycol in a reaction solution by carrying out a DNA amplification and a reaction solution containing tetraethylene glycol for carrying out a DNA amplification are disclosed.07-05-2012
20120171727NOVEL GENOME WALKING METHOD FOR CLONING OF UNKNOWN DNA SEQUENCES ADJACENT TO KNOWN SEQUENCE - A method and a kit for cloning a nucleotide sequence adjacent to a known nucleotide sequence by PCR are disclosed. The method includes 1) preparing a DNA fragment with cohesive ends by cleaving a DNA that contains the known first nucleotide sequence and the second nucleotide sequence adjacent thereto using a restriction enzyme; 2) modifying the 3′ end of the DNA fragment with a nucleotide analogue to block further elongation of the DNA fragment; 3) linking the cohesive ends of the 3′ end-modified DNA fragment with a linker, adapter or cassette having a cohesive end sequence complementary thereto; and 4) performing PCR using a known first nucleotide sequence-specific primer, and a linker, adapter or cassette sequence-specific primer.07-05-2012
20120171725Amplicon Rescue Multiplex Polymerase Chain Reaction for Amplification of Multiple Targets - Disclosed is a method for amplifying and detecting polynucleotides which can provide sensitive, specific detection of multiple targets from a clinical specimen within a relatively short time.07-05-2012
20100047876HIERARCHICAL ASSEMBLY OF POLYNUCLEOTIDES - Methods, compositions and apparatuses for hierarchical assembly of oligonucleotide sequences are provided.02-25-2010
20120178129GENE SYNTHESIS METHOD - The present invention relates to polymerase chain reaction (PCR)-based methods for the one-step synthesis of nucleic acid molecules, wherein the amplification primers used in said methods are designed such that they have two distinct melting temperatures in order to minimize the competition between polymerase cycling assembly (PCA) and polymerase chain reaction (PCR) amplification in the one-step nucleic acid synthesis and to maximize the emerging full-length amplification, as well as kits for use in such methods.07-12-2012
20120178130REUSABLE PCR AMPLIFICATION SYSTEM AND METHOD - A DNA amplification device utilizing a polydimethylsiloxane (PDMS) and silicon substrate coated with spin-on glass (SOG) is provided. This PDMS layer is irreversibly bonded to the SOG layer of the silicon substrate using oxygen plasma. The amplification device is an inexpensive, microfluidic device, which can be utilized as a portable thermo-cycler to perform PCR amplification of DNA in the field.07-12-2012
20100009412Novel Oligonucleotide Primers and Methods for DNA Replication - The present invention relates to methods and oligonucleotide primers for initiating polymerase-catalyzed DNA synthesis at a target region on a polynucleotide template, using an oligonucleotide primer comprising (i) a 5′ universal sequence or extension sequence; (ii) a 3′ target-specific primer sequence corresponding to a target region on the polynucleotide template; and (iii) an extension sequence linking the universal sequence or extension sequence with the target specific primer sequence.01-14-2010
20090061489MICROFLUIDIC DEVICES WITH INTEGRATED RESISTIVE HEATER ELECTRODES INCLUDING SYSTEMS AND METHODS FOR CONTROLLING AND MEASURING THE TEMPERATURES OF SUCH HEATER ELECTRODES - The invention relates to methods and devices for control of an integrated thin-film device with a plurality of microfluidic channels. In one embodiment, the microfluidic device includes a microfluidic chip comprising a first zone having a plurality of microfluidic channels and a second zone having a plurality of microfluidic channels, wherein the microfluidic channels in the first and second zones are in fluid communication. The microfluidic device further comprising a thin-film heater in thermal communication with each of the microfluidic channels in the first and second zones. The microfluidic device also includes a control system configured to independently control the temperature of each of the thin-film heaters using pulse width modulation (PWM) control signals that are optimized for each of the thin-film heaters.03-05-2009
20090061488Method of synthesizing a target polynucleotide encoding a protein - The present invention provides a method of synthesizing a target polynucleotide encoding a protein, which uses a primer extension technique to constitute the target polynucleotide sequence. Preferably, the method is applied in a method for highly expressing a protein encoded by the target polynucleotide in a host.03-05-2009
20100291635CHIMERIC PRIMERS FOR IMPROVED NUCLEIC ACID AMPLIFICATION REACTIONS - Methods are provided for amplification of a nucleic acid sequence. The method use RNA/DNA chimeric oligonucleotides as primers. The primers have RNA residues scattered along their length and no two ribonucleotides in the prime are adjacent to one another. The methods are useful for reducing non-specific amplification products, such as primer dimers. The invention also provides kits comprising RNA/DNA chimeric oligonucleotide primers for practicing the amplification methods.11-18-2010
20120258500PCR REACTION MIXTURES WITH DECREASED NON-SPECIFIC ACTIVITY - The present invention provides methods for improving the specificity of nucleic acid amplification comprising incubating a nucleic acid molecule with a polymerase-Sso7 DNA binding domain conjugate and arginine, spermidine, or spermine. The present invention also provides reaction mixtures and kits for improving the specificity of nucleic acid amplification.10-11-2012
20110124053POLYMERASE INCORPORATION OF NON-STANDARD NUCLEOTIDES - The disclosed invention teaches processes to amplify oligonucleotides by contacting templates and primers with DNA polymerases and triphosphates of non-standard nucleotides, which form nucleobase pairs fitting the standard Watson-Crick geometry, but joined by hydrogen bonding patterns different from those that join standard A:T and G:C pairs. Thus, this invention relates to nucleotide analogs and their derivatives that, when incorporated into DNA and RNA, expand the number of replicatable nucleotides beyond the four found in standard DNA and RNA. The invention further relates to polymerases that incorporate those non-standard nucleotide analogs into oligonucleotide products using the corresponding triphosphate derivatives, and more specifically, polymerases and non-standard nucleoside triphosphates that support the polymerase chain reaction (PCR), including PCR where the products contain more than one non-standard nucleotide unit. Examples are provides that show this process using 6-amino-5-nitro-3-(1′-beta-D-2′-deoxyribofuranosyl)-2(1H)-pyridone to implement the non-standard “small” donor-donor-acceptor (pyDDA) hydrogen bonding pattern, and 2-amino-8-(r-beta-D-2′-deoxyribofuranosyl)-imidazo[1,2-α]-1,3,5-triazin-4(8H)-one to implement the “large” acceptor-acceptor-donor (puADD) pattern.05-26-2011
20110124050METHOD FOR SYNTHESIZING A CDNA IN A SAMPLE IN AN ENZYMATIC REACTION - The present invention relates to a method for synthesizing a cDNA in a sample in an enzymatic reaction, characterized in that the method comprises the steps: simultaneously providing of a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, a buffer, at least one ribonucleotide, at least one deoxyribonucleotide, an anchor oligonucleotide, adding of a sample comprising a ribonucleic acid and incubating the agents from the preceding steps in one or more temperature steps, which are selected so that the first enzyme and the second enzyme display activity, characterized in that additionally an amplification takes place in the same reaction mixture. The invention relates further to a reaction mixture comprising a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, optionally a buffer, optionally at least one ribonucleotide, optionally at least one deoxyribonucleotide, optionally an anchor oligonucleotide and an enzyme with DNA synthesis activity.05-26-2011
20110124049INTEGRATED MICROFLUIDIC DEVICE FOR GENE SYNTHESIS - We report making an integrated micro-fluidic device for synthesizing double stranded DNA from short oligo-nucleotides. We demonstrate successful synthesis of a 760 bp gene segment from a pool of 39 oligonucleotides on a micro-fluidic device using both the one-step and two-step synthesis processes. We also describe purifying the double stranded DNA PCR product and filtering out sequence errors in the double stranded DNA product, all on the same device.05-26-2011
20100221786CYCLIC REVERSE TRANSCRIPTION METHOD - Disclosed is a cyclic reverse transcription method, which comprises performing reverse transcription reaction in one or more cycles, each cycle comprising the steps of: (i) performing reaction with a reverse transcription reaction solution comprising template RNA, RNA-dependent DNA polymerase, a reaction buffer, primers for reverse transcription and dNTPs, and optionally, a stabilizer, at 10° C. to 40° G and, (ii) performing reaction with the resultant reaction solution at 42° to 55° C.09-02-2010
20080280330COVER FOR SAMPLE WITH SAMPLE-SIZE INDEPENDENT HEIGHT ADJUSTMENT - The present invention relates to a device and a method for controlling the temperature of at least one sample. The device comprises at least the following means: (a) means for accommodating at least one sample; (b) means for heating and/or cooling at least one sample; (c) means for covering at least one sample. These means for covering at least one sample comprise at least one movable contact area and first and second means for fixating said at least one movable contact area in at least one defined direction relative to the sample. Therein said first means for fixating matingly engages with a corresponding second means for fixating.11-13-2008
20110039306Reagent Containing a Thermostable Endonuclease - Compositions and methods are provided for preserving the thermostability of an endonuclease in the presence of DNA for uses that include DNA repair. The compositions and methods include the presence of zinc ions.02-17-2011
20110039305THERMOCYCLER AND SAMPLE VESSEL FOR RAPID AMPLIFICATION OF DNA - A thermocycler apparatus and method for rapidly performing the PCR process employs at least two thermoelectric modules which are in substantial spatial opposition with an interior space present between opposing modules. One or multiple sample vessels are placed in between the modules such that the vessels are subjected to temperature cycling by the modules. The sample vessels have a minimal internal dimension that is substantially perpendicular to the modules that facilitates rapid temperature cycling. In embodiments of the invention the sample vessels may be deformable between: a) a shape having a wide mouth to facilitate filling and removing of sample fluids from the vessel, and b) a shape which is thinner for conforming to the sample cavity or interior space between the thermoelectric modules of the thermocycler for more rapid heat transfer.02-17-2011
20110039304Methods to Generate Oligonucleotide Pools and Enrich Target Nucleic Acid Sequences - Methods and compositions for generating oligonucleotide pools are provided. Methods and compositions for enriching target nucleic acid sequences are also provided.02-17-2011
20120322109Covalent Joining of DNA Strands to RNA Strands Catalyzed by Vaccinia Topoisomerase - The present invention provides a method of covalently joining a DNA strand to an RNA strand, a method of tagging a 5′ end of an RNA molecule, a DNA-RNA molecule which has been joined in vitro by the use of a topoisomerase, a method of tagging a 5′ end of an mRNA, and a method of isolating and cloning full-length gene sequences using capped mRNA after subtraction of non-capped RNA.12-20-2012
20120322110Reaction chip, reaction method, temperature controlling unit for gene treating apparatus and gene treating apparatus - The reaction chip of the present invention has a plurality of recesses 12-20-2012
20120322111METHOD FOR AMPLIFYING A TARGET SEQUENCE INCLUDED IN A DOUBLE-STRANDED DNA - the present invention provides an amplification method capable of inhibiting the generating of the undesired amplified double-stranded DNA sequence. In the present method, DNA polymerase, deoxynucleoside triphosphate, the double-stranded DNA, a forward primer, a reverse primer, and a first block nucleic acid are mixed so as to amplify the double-stranded target sequence with use of a polymerase chain reaction. The first block nucleic acid does not serve as an origin for the elongation reaction with the DNA polymerase. The first block nucleic acid is complementary with a part of the third non-amplified sequence which is interposed between the 5′ end and the complimentary single-stranded target sequence. Due to the first block nucleic acid, the generating of the undesired amplified double-stranded DNA sequence is inhibited.12-20-2012
20100203595THERMAL CYCLING APPARATUS AND PROCESS - A thermal cycling apparatus 08-12-2010
20100203596HYBRIDIZATION CHAMBER FOR BIOASSAY AND HYBRIDIZATION METHOD USING THE HYBRIDIZATION CHAMBER - Provided is a hybridization chamber comprising two biochips disposed in one chamber such that surfaces of the biochips, to which biomolecules that are to be analyzed are respectively bonded, face each other, reducing the distance between probe biomolecules, thereby reducing the distance along which biomolecules move in the hybridization chamber. Provided are also methods of hybridizing biomolecules using the hybridization chamber.08-12-2010
20100203597RECOMBINANT REVERSE TRANSCRIPTASES - The present invention relates to a gene that encodes a hyperactive reverse transcriptase having DNA polymerase activity and substantially reduced RNase H activity, vectors containing the gene and host cells transformed with the invention. The present invention also includes a method of producing the hyperactive reverse transcriptase, producing cDNA from mRNA using the reverse transcriptase of the invention, kits and assay templates made using the hyperactive reverse transcriptase.08-12-2010
20110217736SYSTEM FOR HOT-START AMPLIFICATION VIA A MULTIPLE EMULSION - System, including methods, apparatus, compositions, and kits, for making and using compound droplets of a multiple emulsion to supply an amplification reagent, such as a heat-stable DNA polymerase or DNA ligase, to an aqueous phase in which the compound droplets are disposed. The compound droplets may be induced to supply the amplification reagent by heating the multiple emulsion, to achieve hot-start amplification.09-08-2011
20110217735COMPOSITIONS AND METHODS FOR REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION (RT-PCR) - The present invention is directed to compositions and methods useful for the amplification of nucleic acid molecules by reverse transcriptase-polymerase chain reaction (RT-PCR). Specifically, the invention provides compositions and methods for the amplification of nucleic acid molecules in a simplified one- or two-step RT-PCR procedure using combinations of reverse transcriptase and thermostable DNA polymerase enzymes in conjunction with sulfur-containing molecules or acetate-containing molecules (or combinations of such sulfur-containing molecules and acetate-containing molecules), and optionally bovine serum albumin. The invention thus facilitates the rapid and efficient amplification of nucleic acid molecules and the detection and quantitation of RNA molecules. The invention also is useful in the rapid production and amplification of cDNAs which may be used for a variety of industrial, medical and forensic purposes.09-08-2011
20100233762THERMOSTABLE Y-FAMILY POLYMERASES AND CHIMERAS - The present disclosure is related to thermostable Y-family polymerases, in particular several novel Y-family polymerases and chimeras made therefrom, as well as methods of identifying other Y-family polymerases, methods of generating other chimeric Y-family polymerases, methods of amplifying ancient or damaged DNA, and methods of incorporating fluorescent or modified nucleotides into a DNA molecule.09-16-2010
20120276592Microfluidic System and Method for a Polymerase Chain Reaction - A microfluidic system for a polymerase chain reaction is disclosed. The system includes a substrate having three chambers fluidically connected to one another in series, which chambers are held at different temperature levels. An elastic film on the substrate closes the chambers, wherein the chambers connected to one another in series are fluidically closable at the ends of the serial connection. The film above a chamber is movable into the chamber for emptying of the chamber. Thus, without a separate pump, it is possible for a PCR solution to be pumped through the chambers or temperature levels, with the PCR solution in a chamber acquiring the temperature thereof very rapidly.11-01-2012
20100028954Template-dependent nucleic acid polymerization using oligonucleotide triphosphate building blocks - A novel use of a template-dependent polymerase. The novel use is effected by employing the template-dependent polymerase for incorporating at least one oligonucleotide triphosphate onto a nascent oligonucleotide-3′-OH in a template-dependent manner.02-04-2010
20120288897APPARATUS AND METHODS FOR INTEGRATED SAMPLE PREPARATION, REACTION AND DETECTION - An apparatus includes a housing, a reaction vial and a transfer mechanism. The housing defines a first flow path and a second flow path. The housing has transfer port defining an opening in fluid communication with the second flow path and a volume outside of the housing. The transfer port includes a flow control member to limit flow through the opening. The reaction vial is coupled to the housing and defines a reaction volume, which is in fluid communication with the transfer port via the second flow path. The transfer mechanism is configured to transfer a sample from an isolation chamber of an isolation module to the reaction chamber via at least the first flow path when the transfer mechanism is actuated. The transfer mechanism configured to produce a vacuum in the reaction vial to produce a flow of a sample from the isolation chamber to the reaction volume.11-15-2012
20100209970METHOD OF AMPLIFICATION OF GC-RICH DNA TEMPLATES - Methods are provided for increasing the processivity of DNA polymerases on GC-rich templates. The methods relate to providing enhancers and biased ratios of dNTPs, and may be used in DNA amplification reactions. The methods are useful for detecting genotypes associated with GC-rich repeats, including Fragile X Syndrome.08-19-2010
20100190213METHOD AND APPARATUS FOR CONTROLLING THE TEMPERATURE OF REACTION VOLUMES - A method and apparatus for controlling the temperature of one or more reaction volumes contained in respective reaction vessel(s) through absorption of electromagnetic energy by a reactant medium of the reaction volume(s). The method comprises providing one or more-reaction volumes (07-29-2010
20100167353RNASE H-BASED ASSAYS UTILIZING MODIFIED RNA MONOMERS - The present invention pertains to novel oligonucleotide compounds for use in various biological assays, such as nucleic acid amplification, ligation and sequencing reactions. The novel oligonucleotides comprise a ribonucleic acid domain and a blocking group at or near the 3′ end of the oligonucleotide. These compounds offer an added level of specificity previously unseen. Methods for performing nucleic acid amplification, ligation and sequencing are also provided. Additionally, kits containing the oligonucleotides are also disclosed herein.07-01-2010
20100167354METHODS AND COMPOSITIONS FOR AMPLIFICATION OF RNA SEQUENCES - The invention provides methods for isothermal amplification of RNA. The methods are particularly suitable for amplifying a plurality of RNA species in a sample. The methods employ a composite primer, a second primer and strand displacement to generate multiple copies of DNA products comprising sequences complementary to an RNA sequence of interest. In another aspect, the methods employ a single primer (which is a composite primer) and strand displacement to generate multiple copies of DNA products comprising sequences complementary to an RNA sequence of interest. In some embodiments, a transcription step is included to generate multiple copies of sense RNA of an RNA sequence of interest. The methods are useful for preparation of nucleic acid libraries and substrates for analysis of gene expression of cells in biological samples. The invention also provides compositions and kits for practicing the amplification methods, as well as methods which use the amplification products.07-01-2010
20100167355MICROREACTOR WITH VENT CHANNELS FOR REMOVING AIR FROM A REACTION CHAMBER - A microreactor for performing chemical reactions, includes a body (07-01-2010
20130011881METHODS AND COMPOSITIONS FOR MULTIPLEX PCR - The present invention provides methods, compositions, kits, systems and apparatus that are useful for multiplex PCR of one or more nucleic acids present in a sample. In particular, various target-specific primers are provided that allow for the selective amplification of one or more target sequences. In one aspect, the invention relates to target-specific primers useful for the selective amplification of one or more target sequences associated with cancer or inherited disease. In some aspects, amplified target sequences obtained using the disclosed methods, kits, systems and apparatuses can be used in various downstream processes including nucleic acid sequencing and used to detect the presence of genetic variants.01-10-2013
20130011879UNRESTRICTED MUTAGENESIS AND CLONING METHOD - The invention relates to methods for amplifying, modifying, mutating and cloning DNA of any size. These methods comprise a series of PCR reactions, which are punctuated by ligation reactions.01-10-2013
20100129876Pasting Edge Heater - An apparatus and method for thermal cycling including a pasting edge heater. The pasting edge heater can provide substantial temperature uniformity throughout the retaining elements during thermal cycling by a thermoelectric module.05-27-2010
20110159550RNA ISOLATION FROM SOLUBLE URINE FRACTIONS - The invention provides methods for isolating RNA from the soluble fraction of urine. The methods can be used for detecting the presence or absence of an RNA, or quantifying the amount of an RNA. The methods are useful for diagnosing an individual suspected of having a disease by detecting the level of RNA associated with the disease in the soluble fraction of urine. The methods are also useful for prognosing an individual diagnosed with a disease by detecting the level of RNA associated with the disease in the soluble fraction of urine.06-30-2011
20110159549Temperature control for light-emitting diode stabilization - A system is provided that includes a light-emitting diode (LED); a temperature sensor in thermal contact with the LED and capable of measuring an operating temperature and generating an operating temperature signal; and a temperature regulating system capable of receiving the operating temperature signal and regulating the operating temperature based on the operating temperature signal. A method for stabilizing the temperature of an LED is provided. A method is provided that includes providing a system comprising an LED, a reaction region, and a sample in the reaction region; generating excitation beams with the LED; directing excitation beams to the sample; detecting an optical property of the sample to obtain detection data; measuring the- operating temperature of the light emitting diode; and adjusting the detection data of an excitation beam characteristic shift related to the operating temperature, when the LED is operated at the operating temperature to generate the excitation beams.06-30-2011
20110159548METHOD FOR AMPLIFYING MONOMORPHIC-TAILED NUCLEIC ACIDS - The present teachings provide methods for amplifying a plurality of target nucleic acids. In some embodiments, a first oligo-dT-universal primer comprising a 3′ oligo-dT portion and a first 5′ universal portion is used to reverse transcribe a plurality of 3′ poly-A tail-containing nucleic acids. A poly-A tail is added to the 3′ end of the first strand products to form a two-tailed reaction product. The two-tailed reaction product is amplified in a PCR, wherein the PCR comprises the first oligo-dT-universal primer, and a second oligo-dT-universal primer, wherein the second oligo-dT-universal primer comprises a 3′ oligo-dT portion and a second 5′ universal portion, wherein the second 5′ universal portion of the second oligo-dT-universal primer comprises a different sequence than the first 5′ universal portion of the first oligo-dT-universal primer. The present teachings also provide compositions and kits for amplifying target nucleic acids containing monomorphic tails.06-30-2011
20110159547POLYMERASE CHAIN REACTON METHOD, POLYMERASE CHAIN REACTON DROPLET DEVICE, AND POLYMERASE CHAIN REACTON DROPLET DEVICE ARRAY - The present invention discloses a polymerase chain reaction (PCR) method, a PCR droplet device and a PCR droplet device array. The steps of the method comprise that a liquid comprising an analyzer is dropped on the heating coil disposed on the droplet device to form a droplet, then dropping a hydrophobic solution to prevent the droplet from evaporating. When an electric current or a voltage is supplied through at least one conducting wire to heat the heating coil, the inside of the droplet can generate buoyancy to drive the analyzer to move to the top of the inside of the droplet. Subsequently, the analyzer is moved to a periphery of the inside of the droplet so as to form a thermal cycle. Therefore the template is amplified by recycling the thermal cycle.06-30-2011
20130023010METHOD FOR REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION - A method for reverse transcription polymerase chain reaction comprises steps: preparing a capillary, and adding a reverse transcription enzyme into the capillary; and performing a lyophilization process on the RT enzyme contained by the capillary to fabricate the RT enzyme into a lyophilized RT reagent in the capillary. Therefore, a buffer solution, an RNA sample, a polymerase and a primer solution can be added into the capillary to re-dissolve the lyophilized RT reagent and enable a reverse transcription reaction and a polymerase chain reaction of the RNA sample to directly take place inside the capillary, so as to promote convenience and efficiency of experiment.01-24-2013
20130023011METHODS AND COMPOSITIONS FOR MULTIPLEX PCR - The present invention provides methods, compositions, kits, systems and apparatus that are useful for multiplex PCR of one or more nucleic acids present in a sample. In particular, various target-specific primers are provided that allow for the selective amplification of one or more target sequences. In one aspect, the invention relates to target-specific primers useful for the selective amplification of one or more target sequences associated with cancer or inherited disease. In some aspects, amplified target sequences obtained using the disclosed methods, kits, systems and apparatuses can be used in various downstream processes including nucleic acid sequencing and used to detect the presence of genetic variants.01-24-2013
20130171697MICROFLUIDIC DEVICE COMPRISING ROTATABLE DISC-TYPE BODY, AND METHODS OF SEPARATING TARGET MATERIAL AND AMPLIFYING NUCLEIC ACID USING THE SAME - A microfluidic device for controlling a flow of a fluid using centrifugal and rotational force based on a rotatable disc-type body, a method of separating a target material or performing emulsion nucleic acid amplification using the microfluidic device.07-04-2013
20080248535RAPID ONE-STEP REVERSE TRANSCRIPTASE PCR - The present invention is directed to a method for performing a one-step RT-PCR for amplifying a target RNA comprising the steps (i) providing a sample which is supposed to contain said target RNA (ii) adding a reaction mixture comprising all reagents necessary to reverse transcribe said target RNA into cDNA and amplify at least a portion of said cDNA (iii) incubating said sample for a time interval of 0 seconds to 40 seconds at a temperature between 20° C. and 65° C., and (iv) subjecting said sample to multiple cycles of a thermocycling protocol wherein the temperature of said sample is varied between at least a first temperature between 37° C. and 72° C. and a second temperature between 85° C. and 100° C.10-09-2008
20080248534Adaptive Thermal Block Temperature Control Method and System - Aspects of the present teachings describe a method and apparatus for automatically controlling a block temperature to reduce undershooting and overshooting of the temperatures of a sample contained in the block and participating in a polymerase chain reaction (PCR). The adaptive thermal block temperature control begins when a sample temperature enters a sample window region between a preliminary setpoint temperature and a target setpoint temperature for the sample. Based on thermodynamic behavior of the sample and the predetermined phase of PCR, predicting a time period measured subsequent to the preliminary setpoint temperature when the sample will reach the target setpoint suitable for the predetermined phase of PCR. During this time period, varying the block temperature ramp rate with a series of cooling and heating changes to ensure the block temperature reaches the target setpoint temperature at approximately the same time as the sample reaches the same. Synchronizing the block temperature and sample temperature to the target setpoint temperature reduces undershooting and overshooting of the sample temperature and increases the speed and efficiency of the overall PCR process as it relates to the thermal cycling operations.10-09-2008
20080248533 METHOD FOR IN VITRO MOLECULAR EVOLUTION OF PROTEIN FUNCTION - The invention provides a method for generating a polynucleotide sequence or population of sequences from parent single stranded polynucleotide sequences encoding one or more protein motifs, comprising the steps of 10-09-2008
20080241892Modified surfaces for immobilization of active molecules - Modified surfaces, substrates, and methods of producing and using such substrates and surfaces are provided. The substrates and surfaces provide either non-reactive surfaces or low density reactive groups, preferably on an otherwise non-reactive surface, for use in different applications including single molecule analyses.10-02-2008
20080241891Method for the Amplification and Detection of HBV DNA Using a Transcription Based Amplification - The present invention provides a method for the transcription based amplification of a target HBV nucleic acid sequence starting from HBV DNA optionally present in a sample, comprising the steps of, —incubating the sample, suspected to contain HBV, in an amplification buffer with one or more restriction enzymes capable of cleaving the HBV DNA at a selected restriction site, said restriction enzyme creating a defined 3′ end of said HBV DNA stand(s), a promoter-primer, said promoter-primer having a 5′ region comprising the sequence of a promoter recognized by a DNA-dependent RNA polymerase and a 3′ region complementary to the define 3′ end of the DNA strand, a second or reverse primer, having the opposite polarity of the promoter-primer and comprising the 5′ end of the said target sequence, and in case of HBV ssDNA as the target sequence, a restriction primer, maintaining the thus created reaction mixture under the appropriate conditions for a sufficient amount of time for a digestion by the restriction enzyme to take place, subjecting the sample thus obtained to a heat treatment at a temperature and time sufficient to inactivate the restriction enzyme and/or to render at least partially a double strand single stranded, adding the following reagents to the sample: an enzyme having RNA dependent DNA polymerase activity, and maintaining the thus created reaction mixture under the appropriate conditions to a sufficient amount of time for the amplification to take place.10-02-2008
20080241890Pcr Process And Arrangement For Dna Amplification Using Dry Reagents - A PCR process for DNA amplification with thermocyclisation of the corresponding reagents is disclosed, in which total integration of all substances and process steps is achieved in a closed, single-use unit (so-called cartridge) in which the reagents are stored in a storage-stable form at room temperature. According to the process, the water-soluble reagents are covered by a water-insoluble medium, then the DNA to be amplified is supplied and the water-insoluble medium is eliminated, so that the water-soluble reagents are dissolved and PCR can start. In the corresponding arrangement, a test unit designed as a single-use produce (a so-called cartridge) has at least one micro-channel or micro-cavity for receiving a PCR reagent. The PCR reagents in the form of a mixture which can be dried at a negligible vapour pressure and forms a storage-stable substance substance at room temperature adhere to the walls of the micro-channel or micro-cavity and form a thin film covered by an insoluble medium.10-02-2008
20080241893METHODS FOR AMPLIFYING TRICHOMONAS VAGINALIS-DERIVED NUCLEIC ACID - Oligonucleotides useful for determining the presence of 10-02-2008
20080233616Method for Carrying Out the Selective Evolution of Proteins in Vitro - The present invention relates to the production of variants of a protein in an in vitro evolution method, comprising the steps: (A) provision of an in vitro expression system comprising (i) a nucleic acid sequence S which codes for a protein Y which is to be varied, (ii) a target molecule X which is able to bind to the protein Y and/or at least one variant Y′ thereof, (iii) an RNA polymerase (Pol) which is able to transcribe the nucleic acid sequence S, (iv) a reverse transcriptase (RT) which is capable of reverse transcription of transcripts of the nucleic acid sequence S, where either the target molecule X is coupled to Pol and the protein Y is coupled to RT, or the target molecule X is coupled to RT and the protein Y is coupled to Pol, (B)incubation of the in vitro expression system from (A) under conditions which enable transcription, reverse transcription and translation to form variants Y′ of the protein Y and nucleic acid sequences S′ coding therefor, and which favor the formation of variants Y′ with improved binding properties for the target molecule X, (C) isolation and, where appropriate, characterization of those variants Y′ which exhibit improved binding properties for binding to X, and/or isolation of nucleic acid sequence variants S′ coding for Y′.09-25-2008
20080227159Compositions and Methods Using Split Polymerases - The invention relates to compositions and methods utilizing split polymerase enzymes composed of at least two discrete polypeptides that stably associate to form a single polymerase. The invention further relates to nucleic acid constructs for expressing the split polymerases of the invention, and methods for using the split polymerases of the invention. The enzymes of the invention are useful in many applications calling for the detectable labeling of nucleic acids and are particularly useful in quantitative PCR (QPCR) and DNA sequencing applications.09-18-2008
20130143273METHODS FOR NUCLEIC ACID MANIPULATION - A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.06-06-2013
20130143274METHODS FOR NUCLEIC ACID MANIPULATION - A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.06-06-2013
20130177945THERMAL CYCLER FOR PCR INCLUDING TEMPERATURE CONTROL BLADDER - Methods and devices for performing chemical reactions under controlled temperature are described. In one embodiment, the devices provided by the invention comprise a housing diminished hold a reaction chamber disposed within an interior volume of the housing. The reaction chamber has thermally conductive interior and exterior surfaces defining an internal volume therein at a first temperature. The device also includes at least one temperature-control substance at a second temperature into said bladder and expel said temperature-control substance, the bladder expands to about substantially at least a portion of said exterior surfaces of said reaction chamber to enable thermal exchange between said temperature-control substance the said internal volume of reaction chamber.07-11-2013
20130177946Suppression of Non-Specific Amplification with High-Homology Oligonucleotides - The invention comprises suppressor oligonucleotides for reducing amplification of a non-target nucleic acid sequences; the method of designing and using such oligonucleotides, as well as kits and reaction mixtures.07-11-2013
20130137145METHODS FOR NUCLEIC ACID MANIPULATION - A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.05-30-2013
20130137144THERMAL BLOCK WITH BUILT-IN THERMOELECTRIC ELEMENTS - Rapid and uniform temperature changes in the wells of a microplate or any thin-walled plate that contains an array of reaction wells, or in the channels of a multi-channel microfluidics device, are achieved by the use of a thermal block with thermoelectric heating/cooling elements built into the block, or by the use of a thermal block with wedges protruding from its lower surface, with thermoelectric elements placed in surface contact with the angled sides of each wedge.05-30-2013
20110269191THERMAL CYCLER FOR PCR INCLUDING TEMPERATURE CONTROL BLADDER - Methods and devices for performing chemical reactions under controlled temperature are described. In one embodiment, the devices provided by the invention comprise a housing diminished hold a reaction chamber disposed within an interior volume of the housing. The reaction chamber has thermally conductive interior and exterior surfaces defining an internal volume therein at a first temperature. The device also includes at least one temperature-control substance at a second temperature into said bladder and expel said temperature-control substance, the bladder expands to about substantially at least a portion of said exterior surfaces of said reaction chamber to enable thermal exchange between said temperature-control substance the said internal volume of reaction chamber.11-03-2011
20110269193METHOD OF AMPLIFYING A TARGET NUCLEIC ACID BY ROLLING CIRCLE AMPLIFICATION - Provided is a method of amplifying a nucleic acid using rolling cyclic amplification (RCA), including amplifying a nucleic acid together with formation of a single-strand circular DNA template using RCA by reacting a reaction solution including: (a) two hairpin oligos, (b) a target nucleic acid, (c) a DNA ligase, (d) an endonuclease, (e) a DNA polymerase, and (f) a primer.11-03-2011
20130171698HETEROLOGOUS EXPRESSION OF FUNGAL POLYKETIDE SYNTHETIC GENE IN YEAST - The present invention relates to a method of removing an intron contained in a gene from a eukaryotic gene, and linking only the exon sequences to prepare an expression vector comprising the linked sequences. Specifically, the invention relates to a method of preparing an expression vector containing linked exon sequences comprising amplifying exon sequences by PCR as one or more fragments from a giant fungal gene containing an intron, and linking the fragments together with a restriction enzyme-treated vector using the gap repair cloning method; a method of preparing an expression vector containing a full-length cDNA sequence by synthesizing and linking cDNA fragments from a fungal giant gene; a transformant having introduced therein an expression vector prepared by the method; a protein produced by the transformant; and a method of preparing a compound produced by the protein using the expression vector.07-04-2013
20130102034MODIFIED NUCLEOSIDES, NUCLEOTIDES, AND NUCLEIC ACIDS, AND USES THEREOF - The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using thereof.04-25-2013
20130143272THERMAL CYCLER WITH VAPOR CHAMBER FOR RAPID TEMPERATURE CHANGES - Rapid and uniform temperature changes in the wells of a microplate or any thin-walled plate that contains an array of reaction wells or sample receptacles are achieved by the use of heating and cooling elements with a vapor chamber interposed between such elements and the microplate. The upper surface of the vapor chamber and the underside of the sample plate in certain embodiments are complementary in shape, i.e., they have identical but oppositely directed contours in the areas around each of the sample receptacles, to provide continuous surface contact along the surface of each receptacle. In other embodiments, an intermediary plate is placed between the vapor chamber and the well plate, with the top surface of the intermediary plate being complementary in shape to the underside of the well plate.06-06-2013
20110212491REACTION VESSEL - A reaction vessel for carrying out a chemical or biochemical reaction, such as a polymerase chain reaction, said vessel having a coating of parylene or a derivative thereof, on at least the surface which contacts reactants.09-01-2011
20080199916Multiplex targeted amplification using flap nuclease - Methods for multiplex amplification of a plurality of targets of distinct sequence from a complex mixture are disclosed. In one aspect targets are circularized using a single circularization probe that is complementary to two regions in the target that flank a region to be amplified. The targets may hybridize to the circularization probe so that 5′ or 3′ flaps are generated and methods for removing flaps and circularizing the resulting product are disclosed. In another aspect targets are hybridized to dU probes so that 5′ and 3′ flaps are generated. The flaps are cleaved using 5′ or 3′ flap endonucleases or 3′ to 5′ exonucleases. The target sequences are then ligated to common primers, the dU probes digested and the ligated targets amplified.08-21-2008
20130149748DNA POLYMERASES WITH IMPROVED ACTIVITY - Disclosed are DNA polymerases having increased reverse transcriptase efficiency relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.06-13-2013
20130137146METHODS FOR NUCLEIC ACID MANIPULATION - A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.05-30-2013
20130149746DEVICE AND METHOD OF AMPLIFYING NUCLEIC ACIDS BY USING OIL LAYER COMPRISING EXOTHERMAL PARTICLES - The disclosure describes example devices and methods for amplifying nucleic acids by using an oil layer including exothermal particles, forming an emulsion using the oil layer including exothermal particles, and performing a nucleic acids amplification reaction in the emulsion. The nucleic acids amplification reaction may be effectively performed even with a small amount of a sample by maximizing nucleic acids amplification efficiency, reducing reaction time, and improving controls to be convenient.06-13-2013
20130149747DNA POLYMERASES WITH IMPROVED ACTIVITY - Disclosed are DNA polymerases having increased reverse transcriptase efficiency relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.06-13-2013
20110275126Method for Producing Nucleic Acid Sample and Method for Producing Nucleic Acid Amplification Product Using the Same - Provided are a simple method for producing a nucleic acid sample that does not need to use high-risk chaotropic salt in large amounts, does not need to use a limited special carrier, and offers a superior level of safety; and a method for producing a nucleic acid amplification product using the same. With respect to a cell sample containing cells, by releasing nucleic acid complexes from the cells and bringing this treatment liquid into contact with a carrier, the nucleic acid complexes are held in the carrier. Further, in the presence of a dispersion medium, by applying heat treatment to the carrier, DNAs such as genomic DNA and mitochondrial DNA, and RNA are released. Thereby, a nucleic acid sample can be recovered.11-10-2011
20110275125System and Method for Processing a Biological Sample - Systems and methods for processing a biological sample are provided herein. For example, the system can be configured to deaggregate/declump a sample before, during, and/or after sample preparation and/or sample analysis. For example, the system can include a deaggregation device/system in communication with, for example, a nucleic acid amplification process (e.g., an ePCR system). Various embodiments of the deaggregation device are provided herein. For example, in some embodiments, the deaggregation device can include a valve, a valve manifold, a conduit, a channel, or some combinations thereof.11-10-2011
20130122550MULTI-SITE MUTAGENESIS - The present invention provides compositions and improved methods for multi-site directed mutagenesis and DNA shuffling. The present compositions and methods provide increased mutation frequency and increased number of transformants which allow one to sequence only a few clones in order to identify the correct mutants and to obtain the desired mutant by screening large number of transformants in a short time. Moreover, the inclusion of FEN-1, PEF and optimized buffer and cycling conditions provided in the present invention should also facilitate random mutagenized library construction and the mutagenesis of large or difficult templates.05-16-2013
20100291634Annealing control primer and its uses - The present invention relates to an annealing control primer for improving annealing specificity in nucleic acid amplification and its applications to all fields of nucleic acid amplification-involved technology. The present primer comprises (a) a 3′-end portion having a hybridizing nucleotide sequence substantially complementary to a site on a template nucleic acid to hybridize therewith; (b) a 5′-end portion having a pre-selected arbitrary nucleotide sequence; and (c) a regulator portion positioned between said 3′-end portion and said 5′-end portion comprising at least one universal base or non-discriminatory base analog, whereby said regulator portion is capable of regulating an annealing portion of said primer in association with annealing temperature.11-18-2010
20120258501DNA POLYMERASES WITH IMPROVED ACTIVITY - Disclosed are DNA polymerases having increased reverse transcriptase efficiency, mismatch tolerance, extension rate and/or tolerance of RT and polymerase inhibitors relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.10-11-2012
20120258499Recombinase Polymerase Amplification - The present invention features novel, diverse, hybrid and engineered recombinase enzymes, and the utility of such proteins with associated recombination factors for carrying out DNA amplification assays. The present invention also features different recombinase ‘systems’ having distinct biochemical activities in DNA amplification assays, and differing requirements for loading factors, single-stranded DNA binding proteins (SSBs), and the quantity of crowding agent employed.10-11-2012
20100317064Covalent joining of DNA strands to RNA strands catalyzed by vaccinia topoisomerase - The present invention provides a method of covalently joining a DNA strand to an RNA strand comprising (a) forming a topoisomerase-DNA intermediate by incubating a DNA cleavage substrate comprising a topoisomerase cleavage site with a topoisomerase specific for that site, wherein the topoisomerase-DNA intermediate has one or more 5′ single-strand tails; and (b) adding to the topoisomerase-DNA intermediate an acceptor RNA strand complementary to the 5′ single-strand tail under conditions permitting a ligation of the covalently bound DNA strand of the topoisomerase-DNA intermediate to the RNA acceptor strand and dissociation of the topoisomerase, thereby covalently joining the DNA strand to the RNA strand. The present invention also provides a method of tagging a 5′ end of an RNA molecule. The present invention further provides a DNA-RNA molecule which has been joined in vitro by the use of a topoisomerase. The present invention also provides a method of tagging a 5′ end of an mRNA. The present invention provides a method of isolating and cloning full-length gene sequences using capped mRNA after subtraction of non-capped RNA.12-16-2010
20100317063COMPOSITIONS FOR IMPROVED cDNA SYNTHESIS - The present invention generally relates to methods of making cDNA molecules and cDNA libraries. The invention also relates to cDNA molecules and cDNA libraries produced according to these methods, as well as to vectors and host cells containing such cDNA molecules and libraries. The invention also relates to kits for making the cDNA molecules and libraries of the invention.12-16-2010
20100317062HOT START REVERSE TRANSCRIPTION BY PRIMER DESIGN - The present teachings provide methods, compositions, and kits for performing primer extension reactions. In some embodiments, a reverse transcription reaction is performed on a target polynucleotide with a hot start primer comprising a blunt-ended self-complementary stem, and a loop, and extension products form at high temperatures but reduce extension product formation at low temperatures.12-16-2010
20130157315Methods and Apparatuses - A method for amplifying nucleic acid from a higher eukaryotic, such as mammalian or plant, nucleic acid source, the method comprising (a) contacting a sampling device with the source of higher eukaryotic, such as mammalian or plant, nucleic acid such that following said contacting, higher eukaryotic such as mammalian or plant nucleic acid-containing material is adhered to at least part of the sampling device, wherein the sampling device, or part thereof to which the nucleic acid-containing material is adhered, is made of a suitable polymeric material; (b) introducing the sampling device or part thereof to which the nucleic acid-containing material is adhered into a reaction vessel which contains a reaction mixture for carrying out a nucleic acid amplification reaction, without any prior treatment of the nucleic acid-containing material; and (c) performing a nucleic acid amplification reaction.06-20-2013
20130183717CHANNELS WITH CROSS-SECTIONAL THERMAL GRADIENTS - Provided herein are systems, devices, and methods for generating thermal gradients in channels and uses thereof. In particular, provided herein are system, methods, and devices employing first and second thermal layers positioned around a channel in order to create a thermal gradient across a cross-section of the channel having, for example, a nucleic acid denaturation zone, a nucleic acid annealing zone, and a nucleic acid polymerization zone. Such devices find use in, for example, nucleic acid amplification procedures, including digital polymerase chain reaction (dPCR) to temperature cycle droplets for amplification of nucleic acid templates within the droplets.07-18-2013
20110312039DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.12-22-2011
20110312038DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.12-22-2011
20110312037DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.12-22-2011
20110312036MICROCHIP FOR ISOTHERMAL NUCLEIC-ACID AMPLIFICATION REACTION, METHOD FOR PRODUCING THE SAME, AND ISOTHERMAL NUCLEIC-ACID AMPLIFICATION METHOD - A microchip for an isothermal nucleic-acid amplification reaction in which at least one of substances necessary for an isothermal amplification reaction of a nucleic acid is present in a reaction region functioning as a reaction field of the reaction, the at least one of the substances being covered with a thin film that melts at a temperature higher than room temperature and lower than a reaction temperature of the reaction.12-22-2011
20110312035STABILIZED COMPOSITIONS OF THERMOSTABLE DNA POLYMERASE AND ANIONIC OR ZWITTERIONIC DETERGENT - The present invention provides compositions, methods, and kits for protecting thermostable DNA polymerase during amplification reactions conducted at a temperature ranging from about 40° C. to greater than 100° C. The composition comprises a thermostable DNA polymerase and an anionic detergent or zwitterionic detergent.12-22-2011
20130189741COMPOSITIONS AND METHODS FOR REPROGRAMMING MAMMALIAN CELLS - The present invention relates to methods for changing the state of differentiation of a eukaryotic cell, the methods comprising introducing mRNA encoding one or more reprogramming factors into a cell and maintaining the cell under conditions wherein the cell is viable and the mRNA that is introduced into the cell is expressed in sufficient amount and for sufficient time to generate a cell that exhibits a changed state of differentiation compared to the cell into which the mRNA was introduced, and compositions therefor. For example, the present invention provides mRNA molecules and methods for their use to reprogram human somatic cells into pluripotent stem cells.07-25-2013
20120015405DNA POLYMERASES WITH INCREASED 3'-MISMATCH DISCRIMINATION - Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.01-19-2012
20120021463METHOD AND APPARATUS FOR AMPLIFICATION OF NUCLEIC ACID SEQUENCES BY USING THERMAL CONVECTION - The present invention provides a nucleic acid sequence amplification method and apparatuses thereof that are simple in the design and easy to miniaturize and integrate into complex apparatuses, with capability of using DNA polymerases that are not thermostable. In the present invention, a plurality of heat sources are combined to supply or remove heat from specific regions of the sample such that a specific spatial temperature distribution is maintained inside the sample by locating a relatively high temperature region lower in height than a relatively low temperature region.01-26-2012
20120021462RECOMBINASE POLYMERASE AMPLIFICATION - This disclosure describe three related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of the bacterial RecA and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods has the advantage of not requiring thermocycling or thermophilic enzymes. Further, the improved processivity of the disclosed methods allow amplification of DNA up to hundreds of megabases in length.01-26-2012
20120021461ISOTHERMAL STRAND DISPLACEMENT AMPLIFICATION - A method for isothermal DNA amplification comprising: providing to the DNA to be amplified an amplification mix comprising a first primer at least partially complementary to a region of DNA and containing Xanthosine, a second primer at least partially complementary to a region of DNA and containing Xanthosine, a DNA polymerase, an enzyme capable of strand displacement, an enzyme that recognises Xanthosine in double-stranded DNA and causes a nick or excises a base in one DNA strand at or near Xanthosine; and amplifying the DNA substantially without thermal cycling.01-26-2012
20120021460MATERIALS AND METHODS FOR ISOTHERMAL NUCLEIC ACID AMPLIFICATION - A method for isothermal amplification of a target nucleic acid sequence is disclosed. The target nucleic acid is amplified by an enzyme with helicase activity and an enzyme with reverse transcriptase activity and DNA-dependant DNA polymerase activity. Also disclosed is a kit for isothermal amplification of a target nucleic acid sequence, including HPV nucleic acids. The kit comprises a first enzyme with helicase activity and a second enzyme having both reverse transcriptase activity and DNA-dependant DNA polymerase activity.01-26-2012
20120028311CARTRIDGE WITH LYSIS CHAMBER AND DROPLET GENERATOR - Systems, including apparatus, methods, compositions, kits, and software, for preparing, reacting, detecting, and/or analyzing samples in droplet-based assay systems, among others. The disclosure emphasizes, but is not limited to, a disposable cartridge with lysis chamber and droplet chamber, particularly for use in droplet-based assays.02-02-2012
20120028310ISOTHERMAL NUCLEIC ACID AMPLIFICATION METHODS AND COMPOSITIONS - Methods and compositions are provided related to the amplification of target polynucleotide sequences as well as total RNA and total DNA amplification. In some embodiments, the methods and compositions also allow for the immobilization and capture of target polynucleotides with defined 3′ and or 5′ sequences to solid surfaces. The polynucleotides attached to the solid surfaces can be amplified or eluted for downstream processing. In some cases, nucleotides attached to solid surfaces can be used for high throughput sequencing of nucleotide sequences related to target DNA or target RNA.02-02-2012
20130196382DNA POLYMERASES WITH IMPROVED ACTIVITY - Disclosed are DNA polymerases having increased reverse transcriptase efficiency relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.08-01-2013
20090004702CODON SPECIFIC MUTAGENESIS - Materials and Methods are provided for replacing one or more amino acids in a polypeptide with an amino acid of choice to form mutant proteins. Both naturally and non-naturally occurring amino acids can be inserted. A population of mutant proteins can be created in which an amino acid residue has replaced an existing residue at random locations along the primary sequence of the protein. The provided techniques allow for the study of proteins and development of proteins with improved functionalities.01-01-2009
20120058520METHOD FOR SYNTHESIS OF DOUBLE-STRANDED DNA CORRESPONDING TO RNA, AND METHOD FOR AMPLIFICATION OF THE DNA - An object of the present invention is to provide an inexpensive and simple method for synthesis of a double-stranded DNA corresponding to a particular RNA, and a method for amplification of the aforementioned double-stranded DNA. The present invention relates to a method for synthesis of a double-stranded DNA having a nucleotide sequence corresponding to template RNA having polyA, comprising step 1 in which reverse transcription reaction of template RNA is carried out employing oligo(dT)primer to which DNA fragment having a known sequence has been added at the 5′-terminal, to obtain a single-stranded DNA, and step 2 in which double strand formation reaction of single-stranded DNA obtained in step 1 is carried out employing a random primer to which DNA fragment having a known sequence has been added at the 5′-terminal, in the presence of polymerase which does not have 3′→5′ exonuclease activity nor strand displacement activity, to obtain a double-stranded DNA, as well as a method for amplification of a double-stranded DNA having a nucleotide sequence corresponding to template RNA having polyA, further comprising step 3 in which PCR reaction is carried out using the obtained double-stranded DNA.03-08-2012
20130203119NUCLEIC ACID MODIFYING ENZYMES - This invention provides for an improved generation of novel nucleic acid modifying enzymes. The improvement is the fusion of a sequence-non-specific nucleic-acid-binding domain to the enzyme in a manner that enhances the ability of the enzyme to bind and catalytically modify the nucleic acid.08-08-2013
20130203120METHOD FOR CELL LYSIS AND PCR WITHIN THE SAME REACTION CHAMBER - A method for amplification of a target DNA, comprising the steps of (i) transferring a liquid with a first volume comprising at least one or more living cells into a vessel (ii) adding to said vessel a PCR reaction buffer with a second volume, whereas said second volume is at least 2× as large as said first volume (iii) lysing said at least one or more living cells within said vessel by means of incubation for at least 1 Minute at at least 90° C., and (iv) amplifying said target by means of a polymerase chain reaction without performance of an intermediate purification step.08-08-2013
20120088275REAGENTS AND METHODS FOR PCR - Modified double-stranded oligonucleotides that have terminal regions on each of their strands, that have a hybrid length of 6-50 nucleotides long, that have a melting temperature Tm of at least 32° C., and that include 2-4 modifying groups, each covalently attached to a different terminal region, preferably to a terminal nucleotide, said modifying groups being polycyclic substituents that do not have bulky portions that are non-planar, said modified olgonucleotide being capable of binding to the 5′ ex-nuclease domains of DNA polymerases and, when included in a PCR or other primer-dependent DNA amplification reaction at a concentration, generally not more than 2000 nM, that is effective for at least one of the functions of suppressing mispriming, increasing polymerase selectivity against 3′ terminal mismatches, increasing polymerase selectivity against AT-rich 3′ ends, reducing scatter among replicates, suppressing polymerase 5′ exonuclease activity, and inhibiting polymerase activity; as well as amplification reaction mixtures containing such modified double-stranded oligonucleotides, and amplification reactions, amplification assays and kits that include such modified double-stranded oligonucleotides.04-12-2012
20120094332DNA POLYMERASES AND MUTANTS THEREOF - The present invention provides polypeptides having a nucleotide polymerase activity and method of enhancing polymerase activity. The polypeptides of the present invention may possess both a DNA-dependent DNA polymerase activity and an RNA-dependent DNA polymerase activity, i.e., a reverse transcriptase activity. The polypeptides of the present invention may be used in any application including, but not limited to, DNA sequencing reactions, amplification reactions, cDNA synthesis reactions, and combined cDNA synthesis and amplification reactions, e.g., RT-PCR.04-19-2012
20130210078METHODS AND KITS FOR REDUCING NON-SPECIFIC NUCLEIC ACID AMPLIFICATION - Methods and kits for efficient amplification of nucleic acids are provided. The disclosure generally relates to methods and kits for nucleic acid amplification of target nucleic acids of interest. The methods described herein promote the synthesis of the target nucleic acid (i.e., template nucleic acid) by reducing the production of undesirable primer-dimer structures and chimeric nucleic acid products during the amplification process by using novel modified primers.08-15-2013
20130210079REAGENTS AND METHODS FOR AUTOLIGATION CHAIN REACTION - The invention relates to the exponential amplification of specific target nucleic acids. The invention provides methods, reagents and kits for carrying out such exponential amplification via the autoligation chain reaction (ACR).08-15-2013
20130210080PC BOARD-BASED POLYMERASE CHAIN REACTION SYSTEMS, METHODS AND MATERIALS - An apparatus for performing a Polymerase Chain Reaction (PCR) is disclosed. The apparatus comprises a PCR chamber for performing a Polymerase Chain Reaction and a printed circuit board (PCB) fluidic device. The PCR chamber is a fluidic chamber and is located in, or is part of, the printed circuit board (PCB) fluidic device. A method for manufacturing an apparatus for performing the Polymerase Chain Reaction and a method for performing the Polymerase Chain Reaction are further disclosed.08-15-2013
20130210081THERMAL CYCLER AND THERMAL CYCLE METHOD - A thermal cycler (08-15-2013

Patent applications in class Acellular exponential or geometric amplification (e.g., PCR, etc.)

Patent applications in all subclasses Acellular exponential or geometric amplification (e.g., PCR, etc.)