Methods for simultaneous amplification of target loci

  • US 10,538,814 B2
  • Filed: 04/30/2019
  • Issued: 01/21/2020
  • Est. Priority Date: 05/18/2010
  • Status: Active Grant
First Claim
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1. A method for amplifying and sequencing DNA, comprising:

  • ligating adaptors to cell-free DNA isolated from a biological sample, wherein the adaptors each comprises a universal priming site;

    performing a first PCR to simultaneously amplify at least 10 target loci using a universal primer and at least 10 target-specific primers in a single reaction volume;

    performing a second, nested PCR to simultaneously amplify the at least 10 target loci using the universal primer and at least 10 inner target-specific primers in a single reaction volume, wherein at least one of the primers comprises a sequencing tag;

    performing high-throughput sequencing to sequence the amplified DNA comprising the target loci.

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