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Single molecule arrays for genetic and chemical analysis

  • US 8,445,194 B2
  • Filed: 06/13/2006
  • Issued: 05/21/2013
  • Est. Priority Date: 06/15/2005
  • Status: Active Grant
First Claim
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1. A method of identifying a sequence of a target polynucleotide, said method comprising:

  • a) providing a substrate comprising a plurality of discrete regions, wherein a plurality of concatemers are randomly disposed on said plurality of said discrete regions, wherein at least a majority of said discrete regions comprises a single concatemer, wherein at least a majority of said concatemers at said plurality of discrete regions are optically resolvable, andwherein each concatemer comprises a plurality of monomeric units and each monomeric unit comprises;

    i) a first target sequence of said target polynucleotide;

    ii) an adaptor comprising a Type IIs endonuclease restriction site, wherein said first target sequence is adjacent to said adaptor;

    b) applying a first set of probes to said substrate such that one or more probes from said first set hybridizes to said adaptor;

    c) applying a second set of probes to said substrate such that one or more probes from said second set hybridizes to said first target sequence;

    d) ligating probes from said first set and probes from said second set that are hybridized to adjacent sequences of said monomeric unit to form a ligated complex;

    e) detecting said ligated complex thereby identifying a nucleotide of said target polynucleotide;

    f) stripping off said ligated complex;

    g) repeating steps (b) through (f) in order to determine said sequence of said target polynucleotide.

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