Patent application number | Description | Published |
20090117622 | MIXTURE 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 |
20110124050 | METHOD 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 |
20110151459 | SIMULTANEOUS DETECTION OF MULTIPLE NUCLEIC ACID SEQUENCES IN A REACTION - The present invention relates to a method for simultaneously amplifying and detecting nucleic acid sequences in a reaction comprising the following steps: (i) providing a sample comprising at least one nucleic acid molecule; (ii) providing reagents for performing an amplification reaction, wherein the reagents comprise at least four, preferably at least five, more preferably at least six probes, wherein (a) each of the probes is specific for a nucleic acid sequence; (b) at least two, preferably at least three probes carry the same label; and (c) each of the probes that carry the same label has a melting temperature (Tm) which differs by more than 2° C. from the other probes with the same label when they are dissociated from their target nucleic acid sequence by heating; (iii) amplifying the nucleic acid sequences in the reaction; (iv) detecting the amplified nucleic acids by determining whether the labeled probe has bound its nucleic acid sequence; and (v) detecting the temperature at which each given labeled probe dissociates from the nucleic acid sequence to which it has bound. The invention also relates to kits for the use in such a method. | 06-23-2011 |
20120107818 | DETECTION OF MULTIPLE NUCLEIC ACID SEQUENCES IN A REACTION CARTRIDGE - The present invention relates to a method for amplifying and detecting nucleic acid sequences in a reaction cartridge comprising the following steps, (i) providing a sample comprising at least one nucleic acid molecule, (ii) in a first reaction chamber of the cartridge providing reagents for an amplification reaction, (iii) mixing the sample with the amplification reagents, (iv) amplifying the at least one nucleic acid in the first reaction chamber of the cartridge, (v) transferring at least parts of the amplification reaction into a second and third reaction chamber of the cartridge each comprising a probe set, wherein (a) each probe set consists of at least three probes, (b) each of the probes is specific for a nucleic acid sequence, (c) there are at least two probes in each set which carry an identical label, (d) each of the probes in a given probe set that carries an identical label has a melting temperature (T | 05-03-2012 |
20120190026 | METHOD OF NORMALIZED QUANTIFICATION OF RNA - The present invention is related to normalized quantification of RNAs and to the normalization of quantities of RNAs in samples, e.g. mixtures of RNAs. The present invention relates to method for the normalization of the quantity of a RNA to be quantified in a sample to the total quantity of RNA in the sample; or to the total quantity of a specific class of RNA in the sample. | 07-26-2012 |
20120190027 | LIGATION-BASED METHOD OF NORMALIZED QUANTIFICATION OF NUCLEIC ACIDS - The present invention is related to normalized quantification of nucleic acids and to the normalization of quantities of nucleic acids in samples, e.g. mixtures of nucleic acids. The present invention relates to method for the normalization of the quantity of a nucleic acid to be quantified in a sample to the total quantity of nucleic acid in the sample; or to the total quantity of a specific class of nucleic acid in the sample. | 07-26-2012 |
20120202198 | PROCESS FOR THE SYNTHESIS OF A CDNA IN A SAMPLE IN AN ENZYMATIC REACTION - This invention relates to a process for synthesis of a cDNA in a sample, in an enzymatic reaction, whereby the process comprises the steps: simultaneous preparation 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; addition of a sample that comprises a ribonucleic acid; and incubation of the agents of the previous steps in one or more temperature steps, which are selected such that the first enzyme and the second enzyme show activity. The invention further relates to a reaction mixture that comprises 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, and optionally an anchor oligonucleotide. Moreover, the invention relates to a kit that comprises a corresponding reaction mixture. | 08-09-2012 |
20120252014 | METHOD OF NORMALIZED QUANTIFICATION OF NUCLEIC ACIDS USING ANCHOR OLIGONUCLEOTIDES AND ADAPTER OLIGONUCLEOTIDES - The present invention is related to normalized quantification of nucleic acids and to the normalization of quantities of nucleic acids in samples, e.g. mixtures of nucleic acids. The present invention relates to method for the normalization of the quantity of a nucleic acid to be quantified in a sample to the total quantity of nucleic acid in the sample; or to the total quantity of a specific class of nucleic acid in the sample. | 10-04-2012 |
20140057263 | MODIFIED HYBRIDIZATION PROBES - The invention relates to a method for detecting a nucleic acid of an organism in a composition, comprising the steps of, (i) amplifying the nucleic acid to be detected, (ii) during or after amplification, hybridizing to said nucleic acid to be detected a first probe that comprises an abasic site additionally optionally carrying a detectable label, (iii) wherein the position of the abasic site corresponds to a position in said nucleic acid to be detected, known to have a polymorphism in said organism, and wherein said nucleic acid is detected if hybridization occurs. | 02-27-2014 |
20140147843 | METHOD FOR QUANTIFYING HUMAN DNA USING AN INTERNAL CONTROL - The present invention relates to a method for quantifying and/or detecting one or more nucleic acids of a genome in a sample, wherein in an amplification reaction, (i) a first nucleic acid is amplified, the locus that is amplified is a multicopy locus (MCL) within the genome, wherein the locus shares at least 80% sequence identity to a sequence according to SEQ ID NO. 1 over a stretch of 80 base pairs, and wherein the multicopy locus has copies on at least two different chromosomes, (ii) a second nucleic acid that has been added as an internal control (IC) is also amplified, and (iii) the amount of amplification product from the amplification of the first nucleic acid is determined. | 05-29-2014 |
20140234834 | METHOD FOR QUANTIFYING HUMAN DNA - The invention provides for a method for quantifying one or more nucleic acids of a genome in a sample comprising the steps of (a) amplifying a first nucleic acid to be quantified, (b) determining the amount of said first nucleic acid by comparison of the amount of amplification product from said first nucleic acid with at least one amplification product from a second template nucleic acid, (c) wherein said second template nucleic acid was generated using whole genome amplification and wherein the starting concentration of the second template nucleic acid is known. | 08-21-2014 |
20150056624 | METHOD FOR ISOLATING NUCLEIC ACIDS FROM A FOOD SAMPLE - The method for isolating nucleic acids from a food sample comprising the following steps: a) obtaining a food enrichment culture; b) transferring a portion of the food enrichment culture into a reaction vessel thereby providing a food enrichment sample and providing a water-immiscible phase in contact with the food enrichment culture; c) lysing the food enrichment sample to provide a lysed sample; d) isolating nucleic acids from the lysed sample. Food enrichment culture samples are known to contain high concentrations of organic and/or liposoluble inhibitors. By contacting the enrichment culture sample with a water-immiscible phase before the actual DNA extraction procedure starts, part of the lipophilic inhibitors is expected to cross the phase interface due to an enhanced solubility in the organic phase and thereby become depleted. The water-immiscible phase provided according to the invention interacts directly with the sample material throughout bacteria lysis and DNA extraction thereby optimizing the subsequent DNA purification processes due to the depletion of food-borne inhibitors. This method yields nucleic acids, in particular DNA that is of an improved quality and purity to be used in a subsequent PCR reaction to detect pathogen nucleic acids in the isolated nucleic acids. | 02-26-2015 |