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Labelled nucleotides

  • US 8,148,064 B2
  • Filed: 07/19/2010
  • Issued: 04/03/2012
  • Est. Priority Date: 12/04/2001
  • Status: Expired due to Fees
First Claim
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1. A method for detecting the extension of a primer annealed to a target polynucleotide, comprising:

  • (a) providing at least one nucleoside comprising(1) a label attached via a first linker to the nucleoside base, and(2) a protecting group attached via a second linker to the 2′

    or 3′

    oxygen of the nucleoside sugar,wherein the first and second linkers are both cleavable under a single set of conditions;

    (b) extending the annealed primer with the nucleoside only if the nucleoside base is complementary to the base at the corresponding position of the target polynucleotide;

    (c) detecting the presence of the label on the extended primer; and

    (d) incubating the extended primer under the single set of cleavage conditions, whereby both the first and second linkers are cleaved;

    thereby detecting the extension of the primer.

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