Methods for enriching populations of nucleic acid samples
First Claim
1. A method of analyzing nucleic acids using the polymerase chain reaction (PCR), comprising:
- (a) providing a double-stranded population of nucleic acids comprising single-stranded overhangs;
(b) annealing ligate linkers to said population of nucleic acids whereby at least a subset of nucleic acids from said double-stranded population have linkers bound at both ends;
(c) amplifying, said subset of nucleic acids by PCR, thereby analyzing said subset of nucleic acids.
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Abstract
The invention provides several methods for reducing the complexity of a population of nucleic acids prior to performing an analysis of the nucleic acids on a nucleic acid probe array. The methods result in a subset of the initial population enriched for a desired property, or lacking nucleic acids having an undesired properly. The resulting nucleic acids in the subset are then applied to the array for various types of analysis. The methods are particularly useful for analyzing populations having a high decree of complexity, for example, chromosomal-derived DNA, or whole genomic DNA, or mRNA population. In addition, such methods allow for analysis of pooled samples.
70 Citations
15 Claims
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1. A method of analyzing nucleic acids using the polymerase chain reaction (PCR), comprising:
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(a) providing a double-stranded population of nucleic acids comprising single-stranded overhangs;
(b) annealing ligate linkers to said population of nucleic acids whereby at least a subset of nucleic acids from said double-stranded population have linkers bound at both ends;
(c) amplifying, said subset of nucleic acids by PCR, thereby analyzing said subset of nucleic acids. - View Dependent Claims (2, 3, 4, 5, 6)
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7. A method of analyzing a subset of nucleic acids within a nucleic acid population using the polymerase chain reaction (PCR), comprising:
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(a) providing a single-stranded population of nucleic acids;
(b) annealing a double-stranded branch primer to at least a subset of said population of nucleic acids;
(c) extending the primer by DNA polymerase;
(d) repealing steps (a) through (c) at least once to produce a subset of nucleic acids that comprise a partially double-stranded primer sequence at both ends;
(e) amplifying said subset of nucleic acids by PCR, thereby analyzing said subset of nucleic acids. - View Dependent Claims (8)
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9. A method of analyzing a subset of nucleic acids within a nucleic acid population using the polymerase chain reaction (PCR), comprising:
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(a) providing a double-stranded population of nucleic acids that comprise cleaved interrupted palindromic sequences at one or both ends;
(b) annealing an adaptor containing fixed nucleotides onto the said population of nucleic acids thereby producing at least a subset of nucleic acids that comprise an adaptor sequence at both ends;
(c) amplifying said subset of nucleic acids by PCR, thereby analyzing said subset of nucleic acids. - View Dependent Claims (10, 11, 12, 13, 14, 15)
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Specification