TARGETED ENRICHMENT AND AMPLIFICATION OF NUCLEIC ACIDS ON A SUPPORT

摘要

Provided herein is a method of selecting target nucleic acids on a support. The method includes providing a plurality of beads each bead comprising one or more oligonucleotides, providing a support with a plurality of primers with a sequence complementary to at least a portion of the oligonucleotides on the beads, contacting the beads with the support wherein the oligonucleotides on the beads bind to the primers on the support, performing an extension reaction by extending the primers on the support to produce capture oligonucleotides, contacting the support comprising the capture oligonucleotides with the target nucleic acids, and extending the capture oligonucleotides bound to target nucleic acids to produce target extension products comprising a sequence complementary to at least a portion of the target nucleic acids. Optionally, the method further includes amplifying the target extension products.

主权项

1. A method of selecting and amplifying target nucleic acids on a support, the method comprising:
(a) providing a plurality of beads each bead comprising one or more oligonucleotides; (b) providing a support with a plurality of primers with a sequence complementary to at least a portion of the oligonucleotides on the beads; (c) contacting the beads with the support under conditions wherein the oligonucleotides on the beads bind to the primers on the support; and wherein each bead comprises one or more pairs of oligonucleotides comprising forward and reverse primer sequences specific to a target nucleic acid sequence; (d) performing an extension reaction by extending the primers on the support to produce extension products comprising a sequence complementary to at least a portion of the oligonucleotides, wherein the extension products are capture oligonucleotides; (e) contacting the support comprising the capture oligonucleotides with the target nucleic acids, the target nucleic acids potentially comprising sequences complementary to one or more of the capture oligonucleotides; (f) extending the capture oligonucleotides bound to target nucleic acids to produce target extension products comprising a sequence complementary to at least a portion of the target nucleic acids; and (g) amplifying the target extension products.