Patent ID: 8664164
Filing Date: 2014-03-04
Classification: C12Q

Abstract:
1. A method for detecting or enriching for a target deoxyribonucleic acid (DNA) present in a nucleic acid sample, said method comprising: (a) fragmenting a nucleic acid sample to generate nucleic acid fragments including a target fragment containing said target DNA; (b) non-specifically ligating an adaptor sequence to an end of said fragments, wherein the adaptor sequence becomes ligated to an end of a strand of said fragments and wherein the end of said fragments to which said adaptor sequence is ligated is the other end of the same strand to which oligonucleotide B of the probe is ligated in step (e); (c) rendering said fragments, including said target fragment, at least partially single-stranded, wherein the single-stranded portion includes an end portion and wherein the length of said single-stranded portion is sufficient to allow hybridisation of at least part of the single-stranded portion of said target fragment to the probe of step (d), wherein said rendering is done by denaturation or using an exonuclease; (d) contacting the at least partially single-stranded fragments of step (c) with oligonucleotides A and B of a single target-specific nucleic acid probe, wherein: such that said target fragment becomes annealed to said probe through hybridisation to the first target-specific part of oligonucleotide A resulting in only one target-specific probe-binding event per target fragment; (e) directly or indirectly ligating oligonucleotide B of said probe to the part of the single-stranded portion of said target fragment which is hybridised to oligonucleotide A of said probe to produce a probe-target fragment hybrid; and (f) detecting or enriching for said probe-target fragment hybrid.