GENOME-WIDE SCREENING FOR SNPs AND MUTATIONS RELATED TO DISEASE CONDITIONS
First Claim
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1. A method for identifying a genome-wide mutation, comprising:
- (a) providing DNA from two sample pools of a subject, wherein a first sample pool is obtained from an diseased cell and a second sample pool is obtained from a normal cell;
(b) digesting each DNA sample pool to generate at least two DNA fragments;
(c) ligating a short oligonucleotide adapter to each fragment at a site of mismatch cleavage; and
(d) selectively amplifying and identifying the sequence differences between the two sample pools to identify a mutation.
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Abstract
The present invention provides a genome-wide methodology for identifying single nucleotide polymorphisms and mutations related to disease conditions, such as cancer. Specifically, the invention provides methods for detecting genome-wide mutations by successively amplifying sequence differences between two sample populations.
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10 Claims
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1. A method for identifying a genome-wide mutation, comprising:
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(a) providing DNA from two sample pools of a subject, wherein a first sample pool is obtained from an diseased cell and a second sample pool is obtained from a normal cell; (b) digesting each DNA sample pool to generate at least two DNA fragments; (c) ligating a short oligonucleotide adapter to each fragment at a site of mismatch cleavage; and (d) selectively amplifying and identifying the sequence differences between the two sample pools to identify a mutation. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A method for identifying a oncogenic mutation comprising:
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(a) providing DNA from two sample pools of a subject, wherein a first sample pool is obtained from a cancer cell and a second sample pool is obtained from a normal cell; (b) digesting each DNA sample pool to generate DNA fragments; (c) ligating a short oligonucleotide adapter to each fragment at a site of mismatch cleavage; and (d) selectively amplifying and identifying the sequence differences between the two sample pools to identify an oncogenic mutation.
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9. A method for identifying a genome-wide mutation, comprising:
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(a) providing DNA from two sample pools of a subject, wherein a first sample pool is obtained from an diseased cell and a second sample pool is obtained from a normal cell; (b) digesting each DNA sample pool to generate at least two DNA fragments; (c) ligating a short oligonucleotide adapter carrying a recognition site for a type IIS restriction endonuclease to each fragment at a site of mismatch cleavage; (d) digesting all fragments with a type IIS restriction endonuclease and ligating a second short oligonucleotide adapter to the site of IIS restriction endonuclease cleavage; and (e) selectively amplifying and identifying the sequence differences between the two sample pools to identify a mutation.
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10. A method for identifying a genome-wide mutation, comprising:
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(a) providing DNA from two sample pools of a subject, wherein a first sample pool is obtained from an diseased cell and a second sample pool is obtained from a normal cell; (b) digesting each DNA sample pool with Cell to generate at least two DNA fragments; (c) ligating a short oligonucleotide adapter to each fragment at a site of mismatch cleavage; and (d) selectively amplifying and identifying the sequence differences between the two sample pools to identify a mutation.
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Specification