DIRECT CAPTURE, AMPLIFICATION AND SEQUENCING OF TARGET DNA USING IMMOBILISED PRIMERS Russian patent published in 2015 - IPC C12Q1/68 C12N15/11 

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology, in particular, a method of capturing and amplifying a selected sequence and kits for practising the method. The method comprises obtaining a substrate comprising a first population of surface-bound oligonucleotides and a second population of surface-bound oligonucleotides, wherein the members of said first and second populations of surface-bound oligonucleotides are randomly interspersed on the substrate. Hybridising a first member of the first population of surface-bound oligonucleotides to a selection oligonucleotide comprising a region that hybridises with the first member and a region that contains a genomic sequence. Extending the first member of the first population of surface-bound oligonucleotides to produce a duplex, containing a support-bound selection primer that comprises a sequence that is complementary to the genomic sequence. The selection oligonucleotide is removed by denaturation to leave the extended support-bound selection primer. Hybridising the support-bound selection primer to a nucleic acid fragment comprising the genomic sequence. Extending the support-bound selection primer to produce an extension product that contains a sequence, complementary to the flanking sequence, that flanks the genomic sequence. Amplifying the extension product on the substrate, e.g., by bridge PCR using unextended members of the first and second populations of surface-bound oligonucleotides, to produce a PCR product.

EFFECT: target enrichment on the substrate used for sequencing.

19 cl, 18 dwg, 4 tbl, 1 ex