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Methods for simultaneous amplification of target loci

  • US 10,351,906 B2
  • Filed: 10/27/2016
  • Issued: 07/16/2019
  • Est. Priority Date: 04/21/2014
  • Status: Active Grant
First Claim
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1. A method of amplifying target loci in a nucleic acid sample, the method comprising:

  • (a) contacting a sample comprising target loci with a library of at least 50 non-immobilized, non-identical PCR primers that simultaneously hybridize to at least 50 non-identical target loci to produce a reaction mixture; and

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

    wherein an annealing temperature for the PCR conditions is greater than a melting temperature for at least 95% of the at least 50 non-identical PCR primers;

    wherein a length of an annealing step of the PCR conditions is greater than 5 minutes; and

    wherein at least 50 non-identical target loci are simultaneously amplified.

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