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) hybridizing said target with a capture probe;
(c) locking said hybridized capture probe to said target; and
(d) separating a nucleic acid locked to the capture probe from a nucleic acid that is not locked to a capture probe.
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Abstract
Methods for the selective enrichment of nucleic acids.
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Citations
19 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) hybridizing said target with a capture probe; (c) locking said hybridized capture probe to said target; and (d) separating a nucleic acid locked to the capture probe from a nucleic acid that is not locked to a capture probe. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. 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 selector probe hybridizes to said target; (d) joining said selector probe hybridized to said target; and (e) separating a nucleic acid joined to said selector probe from a nucleic acid not joined to a selector probe. - View Dependent Claims (10, 11, 12, 13, 14)
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15. 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 selector probe hybridizes to said target; (d) joining said selector probe hybridized to said target; and (e) separating a nucleic acid joined to said selector probe from a nucleic acid not joined to a selector probe; wherein said target comprises a first capture moiety, and said selector probe comprises a second capture moiety, the method further comprising; contacting said first capture moiety to a first binding moiety, thereby enriching for said target, and contacting said second capture moiety to a second binding moiety, thereby enriching for said selector probe. - View Dependent Claims (16, 17, 18)
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19. A method for normalizing amplified nucleic acids comprising:
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(a) selecting a first population of oligonucleotides capable of hybridizing to a first target, wherein the first population of oligonucleotides comprises oligonucleotides comprising capture moieties and oligonucleotides lacking capture moieties; (b) obtaining a second population of oligonucleotides capable of hybridizing to a second target, wherein the second population of oligonucleotides comprises oligonucleotides comprising capture moieties; (c) amplifying first and second 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, thereby obtaining a population of normalized amplified nucleic acids comprising capture moieties.
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Specification