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Self-assembled single molecule arrays and uses thereof

  • US 8,609,335 B2
  • Filed: 05/02/2011
  • Issued: 12/17/2013
  • Est. Priority Date: 10/07/2005
  • Status: Active Grant
First Claim
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1. A method of determining sequence information for a target polynucleotide, comprising:

  • (a) providing a plurality of concatemers, each concatemer having been obtained by;

    (i) forming a first circular DNA comprising a fragment of said target polynucleotide and a first adaptor, wherein the first adaptor comprises a binding site for a restriction enzyme that cleaves DNA at a cleavage site separated from said binding site by at least six nucleotides;

    (ii) forming a linearized DNA by a process that comprises cleaving the first circular DNA at a site that is internal to the target polynucleotide using a restriction endonuclease that binds to said binding site, whereby the linearized DNA comprises the first adaptor flanked on each side by a portion of said fragment;

    (iii) forming a second circular DNA comprising said linearized DNA and a second adaptor;

    (iv) amplifying the second circular DNA to form said concatemer;

    (b) forming an array of said concatemers bound to a surface; and

    (c) determining target sequences both upstream and downstream from the second adaptor by a process that comprises hybridizing oligonucleotides to the second adaptor in a plurality of the concatemers in the array.

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