Complexity management of genomic DNA
First Claim
1. A method of producing a second nucleic acid sample from a first nucleic acid sample, wherein said second nucleic acid sample is enriched with a collection of adaptor ligated target sequences, the method comprising:
- fragmenting the first nucleic acid sample to produce sample fragments in a selected size range that comprises a collection of target sequences, wherein the fragmenting includes digestion with an enzyme or enzyme combination, wherein the collection of target sequences comprises between 10,000 and 3,000,000 single nucleotide polymorphisms, wherein greater than 80% of the single nucleotide polymorphisms in the collection of target sequences are at least 10 kilo bases and less than 500 kilo bases from another single nucleotide polymorphism, and wherein the average minimum distance between single nucleotide polymorphisms is between 10 kilo bases and 800 kilo bases;
ligating at least one synthetic adaptor to the sample fragments to produce the collection of adaptor ligated target sequences; and
generating the second nucleic acid sample by amplifying the sample fragments, wherein the collection of adaptor ligated target sequences is enriched in the second nucleic acid sample.
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Abstract
The presently claimed invention provides for novel methods and kits for analyzing a collection of target sequences in a nucleic acid sample. A sample is amplified under conditions that enrich for a subset of fragments that includes a collection of target sequences. The invention further provides for analysis of the above sample by hybridization to an array, which may be specifically designed to interrogate the collection of target sequences for particular characteristics, such as, for example, the presence or absence of one or more polymorphisms.
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Citations
31 Claims
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1. A method of producing a second nucleic acid sample from a first nucleic acid sample, wherein said second nucleic acid sample is enriched with a collection of adaptor ligated target sequences, the method comprising:
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fragmenting the first nucleic acid sample to produce sample fragments in a selected size range that comprises a collection of target sequences, wherein the fragmenting includes digestion with an enzyme or enzyme combination, wherein the collection of target sequences comprises between 10,000 and 3,000,000 single nucleotide polymorphisms, wherein greater than 80% of the single nucleotide polymorphisms in the collection of target sequences are at least 10 kilo bases and less than 500 kilo bases from another single nucleotide polymorphism, and wherein the average minimum distance between single nucleotide polymorphisms is between 10 kilo bases and 800 kilo bases; ligating at least one synthetic adaptor to the sample fragments to produce the collection of adaptor ligated target sequences; and generating the second nucleic acid sample by amplifying the sample fragments, wherein the collection of adaptor ligated target sequences is enriched in the second nucleic acid sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31)
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Specification