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Sequential sequencing

  • US 10,619,206 B2
  • Filed: 11/10/2017
  • Issued: 04/14/2020
  • Est. Priority Date: 03/15/2013
  • Status: Active Grant
First Claim
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1. A method, comprising:

  • coupling a nucleic acid template strand to a solid support in a sequencing platform;

    sequencing the template strand using a first primer to produce a first sequencing read, the first primer comprising a 3′

    portion that anneals to a first binding site in the template strand;

    removing the first primer and an extension product from the template strand;

    annealing a second primer to a genomic location in the template strand that is more than 5 nucleotides away from a binding site of the first primer, wherein a 3′

    portion of the second primer is complementary to the genomic location in the template strand; and

    sequencing the template strand using the second primer to produce a second sequencing read, wherein at least one of the sequencing steps comprises sequencing by synthesis.

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