High throughput screening of populations carrying naturally occurring mutations
First Claim
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1. A method of detecting a mutation in one or more target sequences, comprising:
- (a) providing a plurality of libraries of amplification products, wherein each library comprises one or more target sequences amplified from nucleic acid of a member of a population, and wherein each library is tagged with a sample identifier;
(b) performing high throughput sequencing to sequence the amplification products; and
(c) clustering/aligning the sequences of the amplification products without the use of an enzyme that recognizes and cuts single nucleotide sequence mismatches and without performing heteroduplex analysis, and detecting a member of the population carrying the mutation using the sample identifier.
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Abstract
Efficient methods are disclosed for the high throughput identification of mutations in genes in members of mutagenized populations. The methods comprise DNA isolation, pooling, amplification, creation of libraries, high throughput sequencing of libraries, preferably by sequencing-by-synthesis technologies, identification of mutations and identification of the member of the population carrying the mutation and identification of the mutation.
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19 Claims
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1. A method of detecting a mutation in one or more target sequences, comprising:
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(a) providing a plurality of libraries of amplification products, wherein each library comprises one or more target sequences amplified from nucleic acid of a member of a population, and wherein each library is tagged with a sample identifier; (b) performing high throughput sequencing to sequence the amplification products; and (c) clustering/aligning the sequences of the amplification products without the use of an enzyme that recognizes and cuts single nucleotide sequence mismatches and without performing heteroduplex analysis, and detecting a member of the population carrying the mutation using the sample identifier. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification