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Methods for relatvie quantification of specific nucleic acid sequences

  • US 20070148646A1
  • Filed: 04/11/2003
  • Published: 06/28/2007
  • Est. Priority Date: 04/11/2003
  • Status: Active Grant
First Claim
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1. A method of assessing an amount of a known target nucleic acid sequence in a sample comprising:

  • a) co-amplifying said target nucleic acid sequence and a known amount of a known control nucleic acid sequence to produce respective target and control amplicons, wherein said control nucleic acid sequence is different than said target nucleic acid sequence; and

    b) determining relative amounts of said respective amplicons by determining relative quantities of a primer extension reaction using each of said respective amplicons as a template, wherein said primer extension reaction is performed using a sequential dispensation order of individual nucleotides, such that primer extension reactions of said respective target and control amplicons are performed sequentially of each other, and wherein determining relative quantities of a primer extension reaction comprises comparing a quantity of nucleotides incorporated during said primer extension reaction for said target amplicon with a quantity of nucleotides incorporated during said primer extension reaction for said control amplicon, wherein relative amounts of said respective amplicons are proportional to relative quantities of nucleotides incorporated during said primer extension reactions and said amount of said target nucleic acid sequence in said sample is proportional thereto.

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