Methods and systems for nucleic acid amplification

Methods and systems for nucleic acid amplification

  • CN 105,121,663 B
  • Filed: 12/25/2014
  • Issued: 11/13/2020
  • Est. Priority Date: 12/25/2013
  • Status: Active Grant
First Claim
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1. A method of amplifying a target nucleic acid present in a biological sample obtained from a subject, comprising:

  • (a) providing a reaction vessel comprising the biological sample and reagents necessary for performing nucleic acid amplification, the reagents comprising (i) a deoxyribonucleic acid (DNA) polymerase and optionally a reverse transcriptase, and (ii) a primer set for the target nucleic acid, to obtain a reaction mixture, wherein the biological sample is obtained directly from a subject and provided in the reaction vessel without further processing; and

    (b) subjecting the reaction mixture in the reaction vessel to a plurality of series of primer extension reactions to generate amplification products indicative of the presence of the target nucleic acid in the biological sample, each series comprising ten or more of the following cycles;

    (i) incubating the reaction mixture under denaturing conditions characterized by a denaturing temperature and a denaturing duration, followed by (ii) incubating the reaction mixture under extending conditions characterized by an extending temperature and an extending duration, wherein a single series is different from at least one other single series in the plurality of series with respect to the denaturing conditions and/or the extending conditions,and wherein the method is not for use in diagnosing a disease.

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