SELECTIVE ENRICHMENT OF NUCLEIC ACIDS
First Claim
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1. A method for selective enrichment of a nucleic acid, said method comprising the steps of:
- (a) obtaining a population of nucleic acids, wherein at least some of the nucleic acids in said population comprise a target;
(b) contacting said population of nucleic acids with a nickase, thereby producing a population of nicked nucleic acids;
(c) contacting said population of nicked nucleic acids with an exonuclease, thereby generating a nucleic acid having a single-stranded portion, wherein said single-strand portion comprises at least a portion of said target;
(d) contacting a capture probe to said at least a portion of said target, wherein said probe hybridizes to said target; and
(e) separating a nucleic acid hybridized to said capture probe from a nucleic acid not bound to said capture probe.
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Abstract
Methods for the selective enrichment of nucleic acids.
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Citations
68 Claims
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1. A method for selective enrichment of a nucleic acid, said method comprising the steps of:
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(a) obtaining a population of nucleic acids, wherein at least some of the nucleic acids in said population comprise a target; (b) contacting said population of nucleic acids with a nickase, thereby producing a population of nicked nucleic acids; (c) contacting said population of nicked nucleic acids with an exonuclease, thereby generating a nucleic acid having a single-stranded portion, wherein said single-strand portion comprises at least a portion of said target; (d) contacting a capture probe to said at least a portion of said target, wherein said probe hybridizes to said target; and (e) separating a nucleic acid hybridized to said capture probe from a nucleic acid not bound to said capture probe.
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2-17. -17. (canceled)
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18. A method for selective enrichment of a nucleic acid, said method comprising the steps of:
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(a) obtaining a population of nucleic acids, wherein at least some of the nucleic acids in said population comprise a target; (b) hybridizing said target with a capture probe; (c) locking said probe hybridized probe to said target; and (d) separating a nucleic acid locked to a probe from a nucleic acid that is not locked to a probe. - View Dependent Claims (23, 24, 25, 26, 27, 34, 35)
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19. (canceled)
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37. A method for selective enrichment of a nucleic acid, said method comprising the steps of:
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(a) obtaining a population of nucleic acids, wherein at least some of the nucleic acids in said population comprise a target, said target comprising a portion of the 5′
end of a nucleic acid and a portion of the 3′
end of the nucleic acid;(b) obtaining a selector probe comprising a first and second oligonucleotide annealed together, wherein said first oligonucleotide is complementary to at least a portion of said 5′
end of the nucleic acid and complementary to at least a portion of said second oligonucleotide, and said second oligonucleotide is complementary to at least a portion of said 3′
end of the nucleic acid;(c) contacting said selector probe to said target, wherein said probe hybridizes to said target; (d) joining said selector probe to said target; and (e) separating a nucleic acid joined to said selector probe from a nucleic acid not joined to said selector probe. - View Dependent Claims (40, 42, 45, 49, 50)
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52. A method for selective enrichment of a nucleic acid, said method comprising the steps of:
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(a) obtaining a population of single-stranded nucleic acids, wherein at least some of the nucleic acids in said population comprise a target, said target comprising the 5′
end of a nucleic acid and the 3′
end of the nucleic acid;(b) obtaining a selector probe comprising a first and second oligonucleotide annealed together, wherein said first oligonucleotide comprises a 5′
portion complementary to said 3′
end of the nucleic acid, a spacer portion, and a 3′
portion complementary to said 5′
end of the nucleic acid, said second oligonucleotide being complementary to said spacer portion;(c) contacting said selector probe to said target, wherein said probe hybridizes to said target; (d) joining said selector probe to said target; and (e) separating a nucleic acid joined to said selector probe from a nucleic acid not joined to said selector probe. - View Dependent Claims (53, 56, 64, 65)
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67. A method for normalizing amplified nucleic acids comprising:
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(a) selecting a first population of oligonucleotides having a ratio of oligonucleotides comprising capture moieties to oligonucleotides lacking capture moieties for a first population of oligonucleotides; (b) obtaining a second population of oligonucleotides; (c) amplifying target nucleic acids with said first and second populations of oligonucleotides; and (d) separating amplified targets having incorporated oligonucleotide comprising capture moieties from amplified targets lacking incorporated oligonucleotide capture moieties.
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68-71. -71. (canceled)
Specification