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. 2021 Mar 30;12(4):507. doi: 10.3390/genes12040507

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Limit of detection and minimal required input DNA material. (A) The number of detected (1) and absent (0) targets for each tested validated VAF across 3 replicates is shown. Circle size corresponds to the number of called variants (max = 60). Data were fitted with a logistic regression. The LOD (90) was determined at a VAF of 0.18%. (B+C) Comparison of 0.125% VAF reference standards with 5 ng (n = 4) and 20 ng (n = 3) input DNA. (B) Percentage of detected verified mutations for both tested DNA amounts. A predicted 50% detection rate was calculated for a DNA input of 2.5 ng. (C) Shown are the measured average variant allele frequencies of detected validated 0.125% targets. Dotted line indicates the expected VAF of 0.125%. Data represent mean ± SD (n = 3).