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Nucleotides and primers with removable blocking groups

  • US 9,150,896 B2
  • Filed: 03/01/2013
  • Issued: 10/06/2015
  • Est. Priority Date: 09/06/2012
  • Status: Active Grant
First Claim
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1. A method of amplifying nucleic acids comprising:

  • a) providing a nucleic acid template, an amplification enzyme, an oligonucleotide primer and a plurality of nucleotide precursors, wherein the plurality of nucleotide precursors comprises a plurality of modified nucleotides, each modified nucleotide comprising a 3′

    blocking group, wherein the 3′

    blocking group is not thermolabile;

    b) removing the 3′

    blocking group from the plurality of modified nucleotides in the presence of the amplification enzyme to produce a plurality of unblocked nucleotides; and

    c) amplifying the nucleic acid template using the plurality of unblocked nucleotides and the amplification enzyme present during the removing of the 3′

    blocking group from the plurality of modified nucleotides, wherein the amplifying the nucleic acid template comprises two or more cycles of exponential amplification of the nucleic acid template.

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