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Hydroxymethyl linkers for labeling nucleotides

  • US 9,322,050 B2
  • Filed: 07/25/2013
  • Issued: 04/26/2016
  • Est. Priority Date: 04/23/2009
  • Status: Active Grant
First Claim
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1. A method of incorporating labeled nucleotides into nucleic acid, comprising:

  • a) providing a plurality of nucleic acid template molecules, a polymerase, and a plurality of types of nucleotide analogues, wherein each type of nucleotide analogue is labeled with a unique label and contains a removable chemical moiety capping the 3′

    -OH group, wherein the label is attached to the base through a chemically cleavable linker, said linker created via reaction with a hydroxymethyl group;

    b) incorporating a first nucleotide analogue with said polymerase;

    c) detecting the label of the incorporated nucleotide analogue, andd) chemically cleaving said linker of said incorporated nucleotide analogue such that said label is removed from said base wherein after said chemical cleaving of step d) said linker consists of a hydroxymethyl group directly attached to said base.

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