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. 2018 Apr 12;8:5885. doi: 10.1038/s41598-018-24156-x

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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ddPCR Probes Used. (a) An alternative exon exclusion probe (FAM-labeled) and two alternative exon inclusion probes (HEX-labeled) used in alternative splicing assays; the probes have ZEN-Iowa Black as the dual-quencher. An alternative exon (exB) is indicated in red, and it’s flanking exons (exA and exC) in blue and green. Location of primers (F and R) is shown by arrows and ZEN probes are displayed in their binding exons. (b) Dual-quencher probes used in DMPK copy number quantification in cDNA (upper part) and gDNA (lower part). The rs527221 SNP in exon 10 of DMPK was used to distinguish normal-size and mutant alleles. Position of primers (F and R) is shown by arrows and probes are displayed in their binding sites. (c) Representative 2-dimensional scatter plot displaying droplet populations in separate clusters depending on their fluorescence amplitude following ddPCR for CACNA1S exon 29 (DM2 sample). FAM-positive droplets (FAM+/HEX−), HEX-positive (FAM−/HEX+), double-positive (FAM+/HEX+), and negative droplets (FAM−/HEX−) are shown.