HIGH THROUGHPUT SCREENING OF POPULATIONS CARRYING NATURALLY OCCURRING MUTATIONS

摘要

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.

主权项

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) determining the sequences of the amplification products using high throughput sequencing; and (c) detecting a mutation by 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 identifying a member of the population carrying the mutation using the sample identifier.