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PH MEASUREMENT FOR SEQUENCING OF DNA

  • US 20110171655A1
  • Filed: 03/16/2011
  • Published: 07/14/2011
  • Est. Priority Date: 12/20/2006
  • Status: Abandoned Application
First Claim
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1. A method for obtaining sequence information from a single-stranded DNA template, comprising:

  • a) providing a primer region hybridized to the single-stranded DNA template;

    b) placing multiple copies of the single-stranded DNA template, including the provided primer region, in a reaction chamber in a microfluidic device comprising a plurality of reaction chambers wherein the reaction chamber holds less than 0.1 μ

    L of reaction mixture;

    c) adding to the reaction chamber a mixture containing DNA polymerase and a plurality of nucleotides, allowing incorporation of nucleotides by the DNA polymerase;

    d) measuring a pH change in the reaction chamber, said pH change being a result of the incorporation of nucleotides by the DNA polymerase producing a pH change indicative of the incorporation of a complementary nucleotide from the primer region hybridized to the single-stranded DNA template;

    e) removing unbound nucleotides from the reaction chamber; and

    repeating steps c), d), and e) to obtain the sequence information.

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