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Methods and compositions for inhibiting undesired cleaving of labels

  • US 8,623,598 B2
  • Filed: 03/17/2009
  • Issued: 01/07/2014
  • Est. Priority Date: 03/19/2008
  • 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, a cleaving agent, a cleaving agent scavenger comprising a disulfide bond, and a plurality of nucleotide analogues wherein each nucleotide analogue is labeled with a unique label attached through a cleavable disulfide linker and contains a removable chemical moiety capping the 3′

    -OH group;

    b) incorporating a first nucleotide analogue with said polymerase;

    c) detecting the label of the incorporated nucleotide analogue;

    d) removing the chemical moiety of the incorporated nucleotide analogue capping the 3′

    -OH group with said cleaving agent; and

    e) incorporating a second nucleotide analogue in the presence of said cleaving agent scavenger.

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