CPC C12Q 1/6806 (2013.01) [C12N 9/22 (2013.01); C12N 15/10 (2013.01); C12N 15/1093 (2013.01); C12P 19/34 (2013.01); C12Q 1/68 (2013.01); C12Q 1/6883 (2013.01)] | 20 Claims |
1. A method for preparing a target mutant nucleic acid for subsequent enrichment relative to a wild type nucleic acid comprising
subjecting a nucleic acid sample comprising a double-stranded wild type nucleic acid and a double-stranded target nucleic acid suspected of containing a mutation to a condition that destabilizes the double stranded wild type and target mutant nucleic acids;
contacting the destabilized double stranded wild type and target mutant nucleic acids with a pair of oligonucleotide probes, one of which is complementary to the wild type nucleic acid top strand and the other is complementary to the wild type nucleic acid bottom strand, to permit hybridization of the probes to their corresponding sequences on the wild type and target mutant nucleic acids thereby forming complementary wild-type-probe duplexes on top and bottom strands, and partially complementary target mutant-probe duplexes, wherein at least one of the probes overlaps the suspected mutation; and
exposing the complementary wild-type-probe duplexes and the partially complementary target mutant-probe duplexes to a double strand-specific nuclease (DSN),
wherein the DSN preferentially cleaves the complementary wild type-probe duplexes relative to the partially complementary target mutant-probe duplexes.
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