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Dynamic flux nucleic acid sequence amplification

  • US 9,670,531 B2
  • Filed: 04/26/2016
  • Issued: 06/06/2017
  • Est. Priority Date: 03/28/2007
  • Status: Active Grant
First Claim
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1. A real-time dynamic flux method of nucleic acid sequence amplification, comprising:

  • a. combining a pair of forward and reverse oligonucleotide primers with a target nucleic acid sequence to be amplified; and

    b. amplifying the target nucleic acid sequence by thermocycling the pair of forward and reverse oligonucleotide primers and the target nucleic acid sequence,wherein the pair of forward and reverse oligonucleotide primers are designed for amplification of the target nucleic acid sequence to occur within a 15°

    C. temperature range that is defined by the area contained within the overlap of an annealing curve (TA) of the pair of oligonucleotide primers and the denaturation curve (TD) of the target nucleic acid sequence,wherein each forward and reverse oligonucleotide primer has a melting temperature (Tm) within 15°

    C. of the Tm of the target nucleic acid sequence, andwherein thermocycling comprises;

    i. denaturing the target nucleic acid sequence; and

    ii. annealing of the forward and reverse oligonucleotide primers; and

    iii. extension of the target nucleic acid sequence by the forward and reverse oligonucleotide primers,c. simultaneously detecting the amplified target nucleic acid sequence during said amplifying step.

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