Methods for simultaneous amplification of target loci

  • US 10,557,172 B2
  • Filed: 04/30/2019
  • Issued: 02/11/2020
  • Est. Priority Date: 05/18/2010
  • Status: Active Grant
First Claim
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1. A method for amplifying and sequencing DNA, comprising:

  • isolating cell-free DNA from a biological sample and tagging the isolated cell-free DNA, wherein each tagged DNA molecule comprises a molecular barcode;

    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;

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

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