CPC C12N 15/1065 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6827 (2013.01); C12Q 1/6869 (2013.01)] | 23 Claims |
1. A method for identifying a variant of a gene of interest having less than 95% identity to said gene of interest, in a complex sample, said method comprising:
a) preparing DNA from a complex sample comprising a variant of a gene of interest for hybridization thereby forming a prepared sample DNA, the prepared sample DNA comprising said variant of said gene of interest, wherein said gene of interest comprises a plant pest resistance gene of interest, a plant disease resistance gene of interest, and/or a herbicide tolerance gene of interest;
b) mixing said prepared sample DNA with a labeled bait pool comprising polynucleotide sequences complementary to said plant pest resistance gene of interest, plant disease resistance gene of interest, and/or herbicide tolerance gene of interest;
c) hybridizing the prepared sample DNA to said labeled bait pool under conditions that allow for hybridization of a labeled bait in said labeled bait pool with said variant of said plant pest resistance gene of interest, plant disease resistance gene of interest, and/or herbicide tolerance gene of interest to form one or more hybridization complexes,
wherein said variant of said plant pest resistance gene of interest, plant disease resistance gene of interest, and/or herbicide tolerance gene of interest in the hybridization complexes comprises captured DNA;
d) sequencing said captured DNA to determine a sequence read of said variant of said plant pest resistance gene of interest, plant disease resistance gene of interest, and/or herbicide tolerance gene of interest; and
e) aligning said sequence read to a database of known sequences using a sequence alignment program in order to identify said variant of said plant pest resistance gene of interest, plant disease resistance gene of interest, and/or herbicide tolerance gene of interest having less than 95% identity to said plant pest resistance gene of interest, plant disease resistance gene of interest, and/or herbicide tolerance gene of interest.
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