Patent ID: 7932034

Claim:
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, wherein the primer region is: (i) a hairpin formed by self-hybridization of the single-stranded DNA template, or (ii) a single-stranded oligonucleotide that is complementary to a region of the single-stranded DNA template; b) placing multiple copies of the single-stranded DNA template including the provided primer region in reaction chambers in a microfluidic device comprising a plurality of reaction chambers that are thermally isolated from each other, wherein the reaction chambers have a fluid volume of less than 0.1 μL of reaction mixture; c) adding to said reaction chambers containing said copies a mixture containing DNA polymerase and a plurality of a single species of nucleotides; d) removing unincorporated nucleotides; e) measuring a temperature change in said reaction chambers containing said copies of at least 0.003° C., said temperature change being measured by a probe in proximity to the reaction mixture as a result of the incorporation of nucleotides by the DNA polymerase producing a heat increase indicative of the incorporation of a complementary nucleotide into the primer region hybridized to the single-stranded DNA template; and f) repeating steps c), d), and e) to obtain the sequence information.