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Sequence tag directed subassembly of short sequencing reads into long sequencing reads

  • US 8,383,345 B2
  • Filed: 09/14/2009
  • Issued: 02/26/2013
  • Est. Priority Date: 09/12/2008
  • Status: Active Grant
First Claim
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1. A method for preparing a DNA sequencing library, comprising:

  • (a) incorporating at least one first nucleic acid adaptor molecule into at least one member of a target library comprising a plurality of nucleic acid molecules, wherein at least a portion of the first adaptor molecule comprises a first defined sequence;

    (b) amplifying the plurality of nucleic acid molecules to produce an input library comprising a first plurality of amplified DNA molecules, wherein the amplified molecules comprise sequence identical to or complementary to at least a portion of the first adaptor molecule and sequence identical to or complementary to at least a portion of at least one member of the target library;

    (c) fragmenting the input library to produce a plurality of linear DNA fragments having a first end and a second end;

    (d) attaching at least one second nucleic acid adaptor molecule to one or both ends of at least one of the plurality of linear DNA fragments, wherein at least a portion of the second adaptor molecule comprises a second defined sequence; and

    (e) amplifying the plurality of linear DNA fragments to produce a sequencing library comprising a second plurality of amplified DNA molecules, wherein at least one of the plurality of amplified DNA molecules comprises sequence identical to or complementary to at least a portion of the first adaptor molecule, sequence identical to or complementary to at least a portion of the second adaptor molecule, and sequence identical to or complementary to at least a portion of a member of the target library.

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