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Methods and compositions for isolating nucleic acid sequence variants

  • US 7,635,566 B2
  • Filed: 06/27/2008
  • Issued: 12/22/2009
  • Est. Priority Date: 06/29/2007
  • Status: Active Grant
First Claim
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1. A method of sorting nucleic acid molecules according to the identity of their corresponding unique nucleotide tags, said method comprising:

  • (i) contacting a nucleic acid sample comprising at least two nucleic acid molecules each having a unique tag to a capture primer under hybridization conditions, wherein said capture primer is immobilized on a substrate and comprises a sequence complementary to a capture primer binding site in said at least two nucleic acid molecules, whereby said at least two nucleic acid molecules are immobilized;

    (ii) contacting said immobilized at least two nucleic acid molecules to a polymerization primer under hybridization conditions, wherein said polymerization primer comprises a sequence complementary to all of or a region in the unique tag of a first of said at least two nucleic acid molecules, wherein said polymerization primer binds to the same strand of said first nucleic acid molecules as said capture primer at a location that is 5′

    to said capture primer; and

    (iii) isolating said first nucleic acid molecules by contacting said immobilized at least two nucleic acid molecules to a displacing nucleotide polymerase under nucleic acid polymerization conditions, wherein extension of said polymerization primer displaces said first nucleic acid molecules from said immobilized capture primer;

    wherein said at least two uniquely-tagged nucleic acid molecules are sorted according to the identity of their corresponding unique nucleotide tags.

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