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Methods of amplifying and sequencing nucleic acids

  • US 20060040297A1
  • Filed: 08/01/2005
  • Published: 02/23/2006
  • Est. Priority Date: 01/29/2003
  • Status: Active Grant
First Claim
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1. A method for sequencing a nucleic acid, the method comprising:

  • (a) providing a plurality of single-stranded nucleic acid templates disposed within a plurality of cavities on a planar surface, each cavity forming an analyte reaction chamber, wherein the reaction chambers have a center to center spacing of between 20 to 100 μ

    m and the planar surface has at least 10,000 reaction chambers;

    (b) performing a pyrophosphate based sequencing reaction simultaneously on all reaction chambers by annealing an effective amount of a sequencing primer to the nucleic acid templates and extending the sequencing primer with a polymerase and a predetermined nucleotide triphosphate to yield a sequencing product and, if the predetermined nucleotide triphosphate is incorporated onto the 3′

    end of said sequencing primer, a sequencing reaction byproduct; and

    (c) identifying the sequencing reaction byproduct, thereby determining the sequence of the nucleic acid in each reaction chamber.

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