Targeted transposition for use in epigenetic studies

  • US 10,689,643 B2
  • Filed: 05/22/2014
  • Issued: 06/23/2020
  • Est. Priority Date: 11/22/2011
  • Status: Active Grant
First Claim
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1. A method of making a nucleic acid sequence library comprising:

  • a. extracting chromatin from cells to provide a sample containing chromatin;

    b. adding to said sample containing chromatin at least one assembled conjugate comprising a targeting protein covalently conjugated to a stable transposase;

    transposon complex containing a transposase complexed with a transposon cassette, wherein;

    (i) the targeting protein binds a target protein or a target DNA-binding site; and

    (ii) the transposon cassette comprises;

    (1) transposase recognition sequences required for catalysis of a DNA integration reaction;

    (2) one or more oligonucleotide bar code sequences to uniquely identify the conjugated protein; and

    (3) primer sites for DNA amplification;

    c. allowing said at least one conjugate to locate at its/their target proteins and/or target DNA-binding sites in said chromatin;

    d. tagging nucleic acid in said chromatin with said conjugate by inducing an intermolecular reaction between said transposase recognition sequences and said nucleic acid; and

    e. performing PCR amplification of the tagged nucleic acid using the primer sites.

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