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Methods for determining sequence profiles

  • US 10,253,352 B2
  • Filed: 11/17/2016
  • Issued: 04/09/2019
  • Est. Priority Date: 11/17/2015
  • Status: Active Grant
First Claim
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1. A method of detecting a preselected signature in a nucleic acid, comprising:

  • (a) incorporating a subset of nucleic acid base types into a primed nucleic acid to form an extended nucleic acid, wherein the subset of nucleic acid base types is complementary to at least one and no more than three nucleic acid base types in the primed nucleic acid, wherein incorporation of each of the individual nucleic acid base types is not detected among the subset of nucleic acid base types;

    (b) contacting the extended nucleic acid with a polymerase and a nucleotide with a preselected base type different from the nucleic acid base types in the subset of step (a);

    (c) detecting a ternary complex comprising the extended nucleic acid, the polymerase and the nucleotide with the preselected nucleic acid base type thereby detecting the preselected signature, wherein the nucleotide with the preselected base type is prevented from incorporating into the extended nucleic acid during the detecting;

    (d) incorporating, after step (b), the nucleotide with the preselected based type into the extended nucleic acid;

    (e) repeating steps (a) and (b) at least one time after step (d); and

    (f) determining a number of times the preselected nucleic acid base type is detected to determine a number of times the preselected signature occurs.

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