Methods for detection and quantification of analytes in complex mixtures
First Claim
1. A diverse population of labels, comprising about thirty or more unique labels, wherein each of said unique labels is bound to a nucleic acid molecule.
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Abstract
The invention provides a diverse population of uniquely labeled probes, containing about thirty or more target specific nucleic acid probes each attached to a unique label bound to a nucleic acid. Also provided is a method of producing a population of uniquely labeled nucleic acid probes. The method consists of (a) synthesizing a population of target specific nucleic acid probes each having a different specifier; (b) synthesizing a corresponding population of anti-genedigits each having a unique label, the population having a diversity sufficient to uniquely hybridize to genedigits within the specifiers, and (c) hybridizing the populations of target nucleic acid probes to the anti-genedigits, to produce a population in which each of the target specific probes is uniquely labeled. Also provided is a method of detecting a nucleic acid analyte. The method consists of (a) contacting a mixture of nucleic acid analytes under conditions sufficient for hybridization with a plurality of target specific nucleic acid probes each having a different specifier; (b) contacting the mixture under conditions sufficient for hybridization with a corresponding plurality of anti-genedigits each having a unique label, the plurality of anti-genedigits having a diversity sufficient to uniquely hybridize to genedigits within the specifiers, and (c) uniquely detecting a hybridized complex between one or more analytes in the mixture, a target specific probe, and an anti-genedigit.
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Citations
84 Claims
- 1. A diverse population of labels, comprising about thirty or more unique labels, wherein each of said unique labels is bound to a nucleic acid molecule.
- 7. A diverse population of uniquely labeled probes, comprising about thirty or more target specific nucleic acid probes each attached to a unique label bound to a nucleic acid.
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16. A method of producing a population of labels, comprising synthesizing a population of nucleic acids, each of said nucleic acids within said synthesized population having bound a predetermined ratio of at least two different labels, wherein said population of labels has a diversity of predetermined ratios of about 30 or more.
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23. A method of attaching a label to a nucleic acid probe, comprising hybridizing a nucleic acid probe having a genedigit to an anti-genedigit having a label, said genedigit comprising a set of three or more repeat sequences, said anti-genedigit comprising a cognate set of at least two complimentary repeat sequences, wherein said anti-genedigit specifically hybridizes to said genedigit through a sequence having a complexity less than the number of hybridized base pairs.
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34. A method of producing a population of nucleic acid probes, comprising:
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(a) producing a first population of nucleic acids comprising two or more target specific probes each having at least one genedigit, said genedigit having a set of three or more repeated sequences;
(b) producing a second population of nucleic acids comprising an anti-genedigit having a cognate set of at least two complimentary repeated sequences, and (c) hybridizing said first and second populations of nucleic acids to produce a population of target specific probes attached to an anti-genedigit, wherein said anti-genedigit hybridizes to said genedigit through a sequence having a complexity less than the number of hybridized base pairs.
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43. The method of claim 347 wherein said two or more target specific probes further comprise at least one different genedigit.
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49. A method of producing a population of uniquely labeled nucleic acid probes, comprising:
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(a) synthesizing a population of target specific nucleic acid probes each having a different specifier;
(b) synthesizing a corresponding population of anti-genedigits each having a unique label, said population having a diversity sufficient to uniquely hybridize to genedigits within said specifiers, and (c) hybridizing said populations of target nucleic acid probes to said anti-genedigits, to produce a population wherein each of said target specific probes is uniquely labeled.
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50. A method of detecting a nucleic acid analyte, comprising:
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(a) contacting a mixture of nucleic acid analytes with a plurality of target specific probes each attached to a unique label bound to a nucleic acid under conditions sufficient for hybridization of said probes to said target, and (b) measuring a signal from one or more said target specific probes hybridized to an analyte, wherein said signal uniquely identifies the analyte species. - View Dependent Claims (51, 52, 53, 54, 55, 56, 57, 58, 59, 60)
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61. A method of detecting a nucleic acid analyte, comprising:
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(a) contacting a mixture of nucleic acid analytes under conditions sufficient for hybridization with a target specific probe having at least one genedigit, said genedigit having a set of three or more repeated sequences;
(b) contacting said mixture under conditions sufficient for hybridization with an anti-genedigit having a cognate set of at least two complimentary repeated sequences, and (c) detecting a hybridized complex comprising said analyte, target specific probe and said anti-genedigit, wherein said anti-genedigit hybridizes to said genedigit through a sequence having a complexity less than the number of hybridized base pairs. - View Dependent Claims (62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76)
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77. A method of detecting a nucleic acid analyte, comprising:
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(a) contacting a mixture of nucleic acid analytes under conditions sufficient for hybridization with a plurality of target specific nucleic acid probes each having a different specifier;
(b) contacting said mixture under conditions sufficient for hybridization with a corresponding plurality of anti-genedigits each having a unique label, said plurality of anti-genedigits having a diversity sufficient to uniquely hybridize to genedigits within said specifiers, and (c) uniquely detecting a hybridized complex between one or more analytes in said mixture, a target specific probe, and an anti-genedigit. - View Dependent Claims (79, 80, 81, 82, 83, 84)
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78. A nucleic acid labeling kit, comprising a set of genedigits, a set of anti-genedigits and a unique set of labels bound to a nucleic acid.
Specification