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Four-color DNA sequencing by synthesis using cleavable fluorescent nucleotide reversible terminators

  • US 9,528,151 B2
  • Filed: 04/01/2014
  • Issued: 12/27/2016
  • Est. Priority Date: 12/01/2006
  • Status: Active Grant
First Claim
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1. A method, comprising:

  • a) providing a plurality of primer strands bound to nucleic acid templates, a fraction of said plurality of primer strands comprising a free 3′

    -OH and another fraction of said plurality of primer strands comprising an incorporated labeled nucleotide analogue reversible terminator, wherein each reversible terminator comprises a cleavable moiety bound to the 3-oxygen of the deoxyribose, wherein a label is bound via a chemically cleavable linker; and

    b) extending with a polymerase one or more of said primer strands that has a free 3′

    -OH with an unlabeled nucleotide analogue reversible terminator, wherein each unlabeled analogue comprises a cleavable moiety bound to the 3′

    -oxygen of the deoxyribose.

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