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Methods for simultaneous amplification of target loci

  • US 10,316,362 B2
  • Filed: 10/20/2015
  • Issued: 06/11/2019
  • Est. Priority Date: 05/18/2010
  • Status: Active Grant
First Claim
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1. A method of amplifying target loci of DNA molecules in a nucleic acid sample, the method comprising:

  • (a) contacting the sample and a library of at least 1,000 non-immobilized, non-identical primers that hybridize to at least 1,000 non-identical target loci to produce a reaction mixture in which the concentration of each primer is less than 20 nM;

    wherein the average length of the DNA molecules is less than 200 base pairs; and

    wherein a range of melting temperatures of the at least 1000 non-identical primers is less than 10°

    C.;

    (b) subjecting the reaction mixture to PCR conditions to produce amplified products comprising target amplicons;

    wherein a length of an annealing step of the PCR conditions is greater than 10 minutes;

    wherein the length of the target amplicons is less than 100 nucleotides;

    wherein at least 1,000 different target human loci are amplified, and wherein at least 50% of the amplified products comprise target loci.

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