Methods and systems for solution based sequence enrichment

  • US 9,790,543 B2
  • Filed: 09/22/2011
  • Issued: 10/17/2017
  • Est. Priority Date: 10/23/2007
  • Status: Active Grant
First Claim
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1. A method for reducing the genetic complexity of a plurality of nucleic acid molecules, the method comprising, in order, the steps of:

  • a) providing a plurality of support-immobilized oligonucleotide probes defining a complete sequence of at least one genetic locus having a size of at least 100 kb;

    b) amplifying said support-immobilized oligonucleotide probes in solution in the presence of a nucleotide containing a binding moiety to generate a pool of amplification products derived from the support-immobilized oligonucleotide probes, wherein the pool of amplification products contain the binding moiety, and wherein the pool of amplification products are maintained in solution;

    c) contacting under aqueous conditions the pool of amplification products generated in step b) to a sample that comprises a plurality of fragmented nucleic acid molecules, a subset of the plurality of fragmented nucleic acid molecules being target nucleic acid molecules having target nucleic acid sequences;

    d) capturing the target nucleic acid molecules in hybridization complexes with the pool of amplification products, by providing a binding partner for the binding moiety;

    e) separating the hybridization complexes from unbound and non-specifically bound nucleic acids; and

    f) eluting the captured target nucleic acid molecules from the hybridization complexes in an eluate pool having reduced genetic complexity relative to the sample, wherein the eluate pool is enriched for the target nucleic acid molecules.

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