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Isolation of polymerase-nucleic acid complexes

  • US 8,658,364 B2
  • Filed: 03/22/2012
  • Issued: 02/25/2014
  • Est. Priority Date: 03/23/2011
  • Status: Active Grant
First Claim
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1. A method for isolating a polymerase-nucleic acid complex comprising:

  • forming a polymerase-nucleic acid complex by mixing;

    (a) a polymerase enzyme comprising strand displacement activity, and (b) a nucleic acid comprising a double stranded portion comprising a first strand and a complementary second strand;

    initiating nucleic acid synthesis by the polymerase enzyme to produce a nascent strand complementary to the first strand, thereby displacing a portion of the second strand;

    halting or reducing the rate of nucleic acid synthesis;

    hybridizing a hook oligonucleotide to the complex through a capture region on the hook oligonucleotide that is complementary to at least some of the displaced portion of the second strand; and

    isolating the complex using the hook oligonucleotide.

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