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Methods and compositions for whole genome amplification and genotyping

  • US 7,670,810 B2
  • Filed: 06/17/2004
  • Issued: 03/02/2010
  • Est. Priority Date: 06/20/2003
  • Status: Active Grant
First Claim
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1. A method for detecting at least 10,000 typable loci comprising single-nucleotide polymorphisms (SNPs) of genome fragments, comprising:

  • (a) providing a plurality of genome fragments of a whole genome,wherein said providing comprises representationally amplifying a whole, native genome, andwherein said plurality of genome fragments comprises a concentration of at least 1 μ

    g/μ

    l of DNA;

    (b) contacting said plurality of genome fragments with an array of at least 10,000 predetermined nucleic acid probes immobilized to a substrate,wherein said probes comprise a region of at most 100 nucleotides in length that is complementary to typable loci of a genome, andwherein said probes hybridize with genome fragments to form probe-fragment hybrids;

    (c) modifying the probe of a probe-fragment hybrid while hybridized to said genome fragments by adding a detectable nucleotide or nucleotide analog by a polymerase;

    (d) exposing said modified probes to denaturing conditions, thereby removing hybridized genome fragments; and

    (e) detecting said modified probes, thereby detecting at least 10,000 typable loci comprising SNPs.

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