Patent ID: 7163790

Claim:
A method of identifying a polymorphism using an error-correcting assay, the method comprising: (a) contacting a target nucleic acid comprising a query sequence with a probe oligonucleotide in an assay under conditions in which the probe specifically hybridizes to the target nucleic acid, wherein the 3′ nucleotide of the probe is a labeled query nucleotide and the probe is attached to a discrete surface location, wherein the attachment of the probe to the surface location permits the 3′ nucleotide to participate in enzymatic reactions; (b) providing an error-correcting polymerase, wherein the error correcting polymerase is joined to a sequence non-specific nucleic-acid-binding domain that (i) binds to double-stranded nucleic acid, and (ii) enhances the processivity of the polymerase compared to an identical polymerase not having the sequence non-specific nucleic-acid-binding domain fused to it; (c) incubating the assay under conditions in which the probe is extended by the polymerase, wherein the labeled query nucleotide is cleaved from the probe when mismatched with the query sequence; and (d) detecting the amount of probe in the discrete location that has been labeled, wherein the amount of labeled probe in the discrete location is indicative of the presence or absence of the polymorphism.