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. 2018 Oct 12;10:106. doi: 10.1186/s13195-018-0435-2

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© The Author(s). 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 2 — Digital droplet PCR validation of the exome sequencing data. For each patient, a 2D dot plot is shown, reporting the distribution of fluorescence (on the y-axis FAM amplitude, and on the x-axis HEX amplitude). FAM and HEX are the fluorescent dyes for the SEZ6 mutant and SEZ6 wild type, respectively. On the basis of the fluorescence measurements and the droplet distributions, thresholds (pink lines) were set to 5000 for the FAM channel (y-axis) and 3000 for the HEX channel (x-axis). Negative droplets (gray), FAM-positive (blue), HEX-positive (green), and FAM/HEX double-positive (orange) droplets are reported for the four cases and no-template control (NTC) analyzed. Each case represents the sum of independent reactions