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qPCR-HRM assays quantify alleles within DNA mixtures.

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posted on 2015-03-17, 02:45 authored by Mary Lynn Baniecki, Aubrey L. Faust, Stephen F. Schaffner, Daniel J. Park, Kevin Galinsky, Rachel F. Daniels, Elizabeth Hamilton, Marcelo U. Ferreira, Nadira D. Karunaweera, David Serre, Peter A. Zimmerman, Juliana M. Sá, Thomas E. Wellems, Lise Musset, Eric Legrand, Alexandre Melnikov, Daniel E. Neafsey, Sarah K. Volkman, Dyann F. Wirth, Pardis C. Sabeti

The derivative melt graphs show the Tm profiles of a representative assay (12) tested with mixtures of clinical samples known to contain only one allele at this assay position. The ratios of genomic DNA from samples containing reference (C) allele to alternate (T) allele in the mixtures were: (A) 1:10, 1:4, 1:2, 1:1 and (B) 10:1, 4:1, 2:1 1:1. The other 41 assays not shown were tested similarly and all performed comparably to detect mixed allelic samples.

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