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Methods and compositions for improving fidelity in a nucleic acid synthesis reaction

  • US 7,482,120 B2
  • Filed: 01/28/2005
  • Issued: 01/27/2009
  • Est. Priority Date: 01/28/2005
  • Status: Expired due to Fees
First Claim
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1. A method for inhibiting nucleotide misincorporation in a nucleic acid synthesis reaction, the method comprising conducting a polymerization reaction on a nucleic acid duplex comprising a template and a primer in the presence ofa nucleic acid polymerase, a first detectably labeled nucleotide corresponding to a first nucleotide species complementary to the position opposite the incorporation site at the 3′

  • terminus of the primer,a second nucleotide of a different species, andat least one nucleotide derivative, each of the nucleotide derivatives corresponding to a different nucleotide species than the first nucleotide species,wherein each nucleotide derivative comprises a modification that inhibits formation of a phosphodiester bond between the nucleotide derivative and a free 3′

    hydroxyl on a primer nucleotide.

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