Patent ID: 9702002
Date: 2017-07-11
CPC Classifications: C12Q

Claim:
1. A method of selecting and amplifying polynucleotides on a solid support, comprising: a) providing a plurality of amplification oligonucleotides immobilized on a solid support comprising a planar surface or a population of beads; b) hybridizing a population of oligonucleotide probes to a subset of said amplification oligonucleotides, each of said oligonucleotide probes comprising a first portion which is complementary to the amplification oligonucleotides and a second portion which comprises a sequence identical to a selected region of a template polynucleotide; c) performing an extension reaction to extend hybridised amplification oligonucleotides to produce a population of support-bound capture oligonucleotides and a population of unextended amplification oligonucleotides, each capture oligonucleotide in said population comprising a sequence that is complementary to a selected region of a template polynucleotide; d) applying a population of template polynucleotides to the solid support under conditions such that the template polynucleotides selectively hybridise to the support-bound capture oligonucleotides, wherein a portion of the template polynucleotides comprises an adapter sequence identical to the unextended amplification oligonucleotides; e) extending the support-bound capture oligonucleotides that are hybridized to the template polynucleotides, thereby generating extension products, each of the extension products having a portion complementary to the template polynucleotides and a portion complementary to the unextended amplification oligonucleotides; f) amplifying the extension products, wherein the amplifying comprises annealing one or more of the unextended amplification oligonucleotides to one or more of the extension products and extending said one or more unextended amplification oligonucleotides, thereby producing a solid-phase amplification product; and g) sequencing a first strand of the solid-phase amplification product of step f) to obtain at least part of the nucleotide sequence of the template polynucleotide.