Methods and compositions for whole genome amplification and genotyping
First Claim
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1. A method for indicating the presence of single nucleotide polymorphisms (SNPs) of interest in typable loci of a genome, comprising the steps of:
- (a) providing an array of at least 1,000 different immobilized nucleic acid probes that are attached to a surface for flow-through sample analysis, wherein said at least 1,000 different immobilized nucleic acid probes comprise a region of at most 100 nucleotides in length that are complementary to at least 1,000 different typable loci of a genome;
(b) contacting said array with a population of genome fragments representing 1.7 Gigabases or more of genomic sequence under conditions wherein probe-fragment hybrids are formed;
(c) contacting said probe-fragment hybrids with a polymerase and a nucleotide analog comprising a label under conditions whereby if the typable locus of a fragment contains a SNP of interest, the hybridized probe is modified by incorporation of the nucleotide analog to obtain a labeled probe; and
(d) detecting the labeled probe, thereby indicating the presence of said single nucleotide polymorphisms of interest at typable loci of said genome.
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Abstract
This invention provides methods of amplifying genomic DNA to obtain an amplified representative population of genome fragments. Methods are further provided for obtaining amplified genomic DNA representations of a desired complexity. The invention further provides methods for simultaneously detecting large numbers of typable loci for an amplified representative population of genome fragments. Accordingly the methods can be used to genotype individuals on a genome-wide scale.
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30 Claims
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1. A method for indicating the presence of single nucleotide polymorphisms (SNPs) of interest in typable loci of a genome, comprising the steps of:
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(a) providing an array of at least 1,000 different immobilized nucleic acid probes that are attached to a surface for flow-through sample analysis, wherein said at least 1,000 different immobilized nucleic acid probes comprise a region of at most 100 nucleotides in length that are complementary to at least 1,000 different typable loci of a genome; (b) contacting said array with a population of genome fragments representing 1.7 Gigabases or more of genomic sequence under conditions wherein probe-fragment hybrids are formed; (c) contacting said probe-fragment hybrids with a polymerase and a nucleotide analog comprising a label under conditions whereby if the typable locus of a fragment contains a SNP of interest, the hybridized probe is modified by incorporation of the nucleotide analog to obtain a labeled probe; and (d) detecting the labeled probe, thereby indicating the presence of said single nucleotide polymorphisms of interest at typable loci of said genome. - 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)
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Specification