Nucleic acid capture moieties
First Claim
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1. A method of promoting the hybridization of a target single-stranded nucleic acid and a nucleic acid capture moiety comprising:
- a) providing a unimolecular nucleic acid capture moiety having a double-stranded region with a terminal base and an adjacent single-stranded region, the terminal base of the double-stranded region being all or part of a duplex-binding-ligand binding site, and said single-stranded region being capable of hybridizing to said target nucleic acid, wherein said unimolecular nucleic acid capture moiety is bound to a solid support;
b) forming a reaction mixture comprising said target single-stranded nucleic acid, said nucleic acid capture moiety, and a duplex-binding ligand, under conditions such that the target single-stranded nucleic acid and the nucleic acid capture moiety can hybridize; and
c) allowing said target single-stranded nucleic acid to hybridize to said nucleic acid capture moiety such that said duplex-binding ligand binds to the intermolecular duplex thereby formed,such that the hybridization of the target single-stranded nucleic acid and the nucleic acid capture moiety is promoted.
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Abstract
Nucleic acid capture moieties, methods of using nucleic acid capture moieties, reaction mixtures including nucleic acid capture moieties, and kits including nucleic acid capture moieties are disclosed.
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Citations
24 Claims
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1. A method of promoting the hybridization of a target single-stranded nucleic acid and a nucleic acid capture moiety comprising:
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a) providing a unimolecular nucleic acid capture moiety having a double-stranded region with a terminal base and an adjacent single-stranded region, the terminal base of the double-stranded region being all or part of a duplex-binding-ligand binding site, and said single-stranded region being capable of hybridizing to said target nucleic acid, wherein said unimolecular nucleic acid capture moiety is bound to a solid support; b) forming a reaction mixture comprising said target single-stranded nucleic acid, said nucleic acid capture moiety, and a duplex-binding ligand, under conditions such that the target single-stranded nucleic acid and the nucleic acid capture moiety can hybridize; and c) allowing said target single-stranded nucleic acid to hybridize to said nucleic acid capture moiety such that said duplex-binding ligand binds to the intermolecular duplex thereby formed, such that the hybridization of the target single-stranded nucleic acid and the nucleic acid capture moiety is promoted. - View Dependent Claims (2, 3, 4)
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5. A method of detecting a target single-stranded nucleic acid sequence, comprising:
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a) providing a nucleic acid capture moiety comprising a structure A-B-C-D wherein A is a nucleic acid sequence; B and D are nucleic acid sequences which are capable of hybridizing to each other to form an intramolecular duplex; and C is a linker which covalently links B and D; wherein A is substantially complementary to a target single-stranded nucleic acid sequence and the nucleic acid capture moiety is immobilized on an insoluble support; b) forming a reaction mixture comprising said target single-stranded nucleic acid and said nucleic acid capture moiety under conditions such that a B;
D intramolecular duplex can form and the target single-stranded nucleic acid and the nucleic acid capture moiety can hybridize; andc) detecting the presence or absence of hybridization of the target nucleic acid and the nucleic acid capture moiety; such that the target single-stranded nucleic acid is detected. - View Dependent Claims (6, 7, 8, 9, 10, 11)
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12. A method of detecting a target single-stranded nucleic acid sequence, comprising:
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a) providing a nucleic acid capture moiety comprising a structure A-B-C-D wherein A is a nucleic acid sequence; B and D are nucleic acid sequences which are capable of hybridizing to each other to form an intramolecular duplex; and C is a linker which covalently links B and D; wherein A is substantially complementary to a target single-stranded nucleic acid sequence and the nucleic acid capture moiety is immobilized on an insoluble support; b) forming a reaction mixture comprising said target single-stranded nucleic acid and said nucleic acid capture moiety under conditions such that a B;
D intramolecular duplex can form and the target single-stranded nucleic acid and the nucleic acid capture moiety can hybridize to form an intermolecular duplex, wherein the intermolecular duplex comprises a duplex-binding-ligand binding site; andc) detecting the presence or absence of hybridization of the target nucleic acid and the nucleic acid capture moiety; such that the target single-stranded nucleic acid is detected. - View Dependent Claims (13, 14, 15, 16, 17, 18)
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19. A method of promoting the hybridization of a target single-stranded nucleic acid and a nucleic acid capture moiety comprising:
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a) providing a unimolecular nucleic acid capture moiety having a double-stranded region and an adjacent single-stranded region, said single-stranded region being capable of hybridizing to said target nucleic acid, wherein said unimolecular nucleic acid capture moiety is bound to a solid support; b) forming a reaction mixture comprising said target single-stranded nucleic acid, said nucleic acid capture moiety, and a duplex-binding ligand, under conditions such that the target single-stranded nucleic acid and the nucleic acid capture moiety can hybridize; and c) allowing said target single-stranded nucleic acid to hybridize to said nucleic acid capture moiety to form an intermolecular duplex, wherein the intermolecular duplex comprises a duplex-binding-ligand binding site, such that said duplex-binding ligand binds to said duplex-binding-ligand binding site, such that the hybridization of the target single-stranded nucleic acid and the nucleic acid capture moiety is promoted. - View Dependent Claims (20, 21, 22, 23)
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24. A method of capturing a target single-stranded nucleic acid sequence from a sample, comprising:
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a) providing a nucleic acid capture moiety comprising a structure A-B-C-D wherein A is a nucleic acid sequence; B and D are nucleic acid sequences which are capable of hybridizing to each other to form an intramolecular duplex; and C is a linker which covalently links B and D; wherein A is substantially complementary to the target single-stranded nucleic acid sequence and the nucleic acid capture moiety is immobilized on an insoluble support; b) forming a reaction mixture comprising said target single-stranded nucleic acid and said nucleic acid capture moiety under conditions such that a B;
D intramolecular duplex can form and the target single-stranded nucleic acid hybridizes to said nucleic acid capture moiety; andc) separating the target single-stranded nucleic acid hybridized to said nucleic acid capture moiety from the reaction mixture; such that the target single-stranded nucleic acid is captured.
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Specification