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Compositions and methods for selection of nucleic acids

  • US 9,528,107 B2
  • Filed: 10/31/2013
  • Issued: 12/27/2016
  • Est. Priority Date: 01/31/2012
  • Status: Active Grant
First Claim
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1. A method for enrichment of a target region in a nucleic acid sample comprising:

  • a) digesting the nucleic acid sample with a restriction enzyme that cuts a defined distance from its recognition site to produce a population of double-stranded nucleic acid fragments, wherein fragments containing the target region have known single-stranded overhangs on each end, each overhang being different;

    b) ligating two types of stem-loop adapters to the population of nucleic acid fragments, wherein one type of stem-loop adapter has a single-stranded overhang sequence complementary to a first of the known single-strand overhangs at one end of the fragment comprising the target region, and the other type of stem-loop adapter has a single-stranded overhang sequence complementary to a second of the known single-stranded overhangs on the other end of the fragment comprising the target region;

    c) treating the sample with one or more exonucleases to digest the double-stranded nucleic acid fragments that have one or no stem-loop adapter linked thereto, thus enriching for the target region in the nucleic acid sample.

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