Selecting tag nucleic acids
First Claim
1. A method of selecting a set of tag nucleic acids with minimal cross hybridization to a nucleic acid, said method comprising providing a list of tag nucleic acids, and excluding nucleic acids from the list of tag nucleic acids which cross hybridize to a single complementary nucleic acid under stringent conditions, thereby providing a set of selected tag nucleic acids with minimal cross hybridization to the nucleic acid.
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Abstract
Methods of selecting tag nucleic acids and VLSIPS™ arrays and the arrays made by the methods are used to label and track compositions, including cells and viruses, e.g., in libraries of cells or viruses. In addition to providing a way of tracking compositions in mixtures, the tags facilitate analysis of cell and viral phenotypes.
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Citations
57 Claims
- 1. A method of selecting a set of tag nucleic acids with minimal cross hybridization to a nucleic acid, said method comprising providing a list of tag nucleic acids, and excluding nucleic acids from the list of tag nucleic acids which cross hybridize to a single complementary nucleic acid under stringent conditions, thereby providing a set of selected tag nucleic acids with minimal cross hybridization to the nucleic acid.
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21. A composition comprising a set of tag nucleic acids, which set of tag nucleic acids comprises a plurality of tag nucleic acids, which tag nucleic acids comprise a variable region;
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which variable region for each tag nucleic acid in the set of tag nucleic acids has the same Tm, the same G+C to A+T ratio, the same length and does not cross-hybridize to a probe nucleic acid; and
,wherein the tag nucleic acids in the set of tag nucleic acids cannot be aligned with less than two differences between any two of the tag nucleic acids in the set of tag nucleic acids. - View Dependent Claims (22, 23, 24)
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- 25. A method of labeling a composition, comprising associating a tag nucleic acid with the composition, wherein the tag nucleic acid is selected from a group of tag nucleic acids which do not cross-hybridize and which have a substantially similar Tm.
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31. A method of pre-selecting experimental probes in an oligonucleotide probe array, wherein the probes have substantially uniform hybridization properties and do not cross hybridize, comprising:
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selecting a ratio of G+C to A+T nucleotides shared by the experimental probes in the array;
determining all possible 4 nucleotide subsequences for variable nucleic acids in the probes of the array; and
excluding all probes from the array which contain prohibited 4 nucleotide sub-sequences, wherein 4 nucleotide subsequences are prohibited when the nucleotide subsequences are selected from the group consisting of self-complementary probes, A4 probes, T4 probes, [G,C]4 probes, and probes complementary to constant region sub-sequences. - View Dependent Claims (32, 33)
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34. A method of detecting a plurality of nucleic acids in a sample, comprising
(i) providing an array of experimental oligonucleotide probes, which probes do not cross hybridize under stringent conditions, wherein the ratio of G+C bases in each probe is substantially identical; -
wherein the probes of the array are arranged into probe sets in which each probe set comprises a homogeneous population of oligonucleotide probes;
(ii) hybridizing said array of oligonucleotides to the sample under stringent hybridization conditions; and
(iii) detecting hybridization of the nucleic acids to the array of oligonucleotide probes. - View Dependent Claims (35, 36)
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37. An array of oligonucleotide probes comprising a plurality of experimental oligonucleotide probe sets attached to a solid substrate, wherein
each experimental oligonucleotide probe set in the array hybridizes to a different target nucleic acid under stringent hybridization conditions; -
each oligonucleotide probe in the probe sets of the array comprises a variable region; and
whereinthe nucleic acid probes do not cross-hybridize in the array. - View Dependent Claims (38, 39, 40, 41, 42, 43)
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44. A method of detecting a target nucleic acid comprising providing a population of nucleic acids to an array of oligonucleotide probes and monitoring hybridization of the test nucleic acids to the probes in the array, wherein the array of oligonucleotide probes comprises a plurality of experimental oligonucleotide probe sets attached to a solid substrate, wherein
each experimental oligonucleotide probe set in the array hybridizes to a different target nucleic acid under stringent hybridization conditions; -
each oligonucleotide probe in the probe sets of the array comprises variable region; and
whereinthe nucleic acid probes do not cross-hybridize in the array. - View Dependent Claims (45, 46)
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47. A plurality of recombinant cells comprising tag nucleic acids selected from a set of tag nucleic acids, which set of tag nucleic acids comprises a plurality of tag nucleic acids, which tag nucleic acids comprise a variable region;
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which variable region for each tag nucleic acid in the set of tag nucleic acids has the same Tm, the same G+C to A+T ratio, the same length and does not cross-hybridize; and
,wherein the tag nucleic acids in the set of tag nucleic acids cannot be aligned with less than two differences between any two of the tag nucleic acids in the set of tag nucleic acids. - View Dependent Claims (48, 49, 50, 51, 52)
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53. A kit comprising an array of oligonucleotides, wherein
the array of oligonucleotide probes comprises a plurality of experimental oligonucleotide probe sets attached to a solid substrate; -
each experimental oligonucleotide probe set in the array hybridizes to a different target nucleic acid under stringent hybridization conditions;
each oligonucleotide probe in the probe sets of the array comprises a variable region; and
the nucleic acid probes do not cross-hybridize in the array. - View Dependent Claims (54, 55, 56, 57)
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Specification