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Methods and compositions for targeted nucleic acid sequencing

  • US 10,100,355 B2
  • Filed: 12/05/2014
  • Issued: 10/16/2018
  • Est. Priority Date: 12/09/2013
  • Status: Active Grant
First Claim
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1. A method for preparing a substrate-bound anchor probe comprising a sequence of a target nucleic acid comprising:

  • (a) providing a nucleic acid library comprising a plurality of different nucleic acids, wherein the nucleic acid library comprises a double-stranded target nucleic acid;

    (b) hybridizing a first adapter probe with the target nucleic acid in the presence of a recombinase,wherein the first adapter probe comprises a first portion complementary to a substrate probe and a second portion complementary to at least a portion of the double-stranded target nucleic acid, andwherein the recombinase promotes hybridizing the second portion with the double-stranded target nucleic acid;

    (c) extending the hybridized first adapter probe;

    (d) hybridizing the extended adapter probe with the substrate probe, wherein the substrate probe is immobilized on a first substrate; and

    (e) extending the hybridized substrate probe to form a substrate-bound anchor probe comprising the sequence of the target nucleic acid.

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