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Single nucleotide amplification and detection by polymerase

  • US 7,033,762 B2
  • Filed: 04/01/2002
  • Issued: 04/25/2006
  • Est. Priority Date: 08/29/2001
  • Status: Expired due to Term
First Claim
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1. A method of characterizing a nucleic acid sample comprising the steps of:

  • (a) conducting a DNA polymerase reaction, said reaction comprising the reaction of a template, a non-hydrolyzable primer, at least one terminal phosphate-labeled nucleotide, DNA polymerase and an enzyme having 3′



    5′

    exonuclease activity, wherein said enzyme may be selected from the group consisting of DNA polymerases, exonucleases and combinations thereof, which reaction results in the production of labeled polyphosphate;

    (b) permitting said labeled polyphosphate to react with a phosphatase to produce a detectable species characteristic of the sample;

    (c) detecting said detectable species; and

    (d) characterizing said nucleic acid sample based on said detection.

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