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Methods and apparatus that increase sequencing-by-binding efficiency

  • US 9,951,385 B1
  • Filed: 09/22/2017
  • Issued: 04/24/2018
  • Est. Priority Date: 04/25/2017
  • Status: Active Grant
First Claim
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1. A method of nucleic acid detection, comprising steps of:

  • (a) contacting a primed template nucleic acid with a polymerase and a first mixture of nucleotides under conditions for forming a ternary complex at a nucleotide position in the template, wherein the first mixture comprisesa nucleotide cognate of a first base type comprising a label that produces a first signal;

    a nucleotide cognate of a second base type comprising a label that produces the first signal;

    a nucleotide cognate of a third base type comprising a label that produces a second signal; and

    a nucleotide cognate of a fourth base type comprising a label that produces the second signal;

    (b) contacting the primed template nucleic acid with a polymerase and a second mixture of nucleotides under conditions for forming a ternary complex at the nucleotide position in the template, wherein the second mixture comprisesa nucleotide cognate of the first base type comprising a label that produces the first signal;

    a nucleotide cognate of a second base type comprising a label that produces the second signal;

    a nucleotide cognate of a third base type comprising a label that produces the second signal; and

    a nucleotide cognate of the fourth base type comprising a label that produces the first signal; and

    (c) examining products of steps (a) and (b) for signals produced by a ternary complex that comprises the primed template nucleic acid, a polymerase and a next correct nucleotide.

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