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. 2022 Mar 28;35(8):1022–1033. doi: 10.1038/s41379-022-01058-y

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© The Author(s) 2022

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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Fig. 1 — A Primers and (B) probes were designed to anneal to exons of target genes commonly involved in somatic translocations, including EWSR1, FLI1, ERG, FUS, WT1, PAX3, PAX7, FOXO1, BCOR, CCNB3. C The multiplexed ddPCR amplifications are split into two independent reactions labeled PCR1 and PCR2. Each reaction uses the same complete primer set but only a specific subset of probes such that the two PCR reactions produce a unique combination of fluorescent signals (characterized by either FAM, HEX, or both) that is characteristic of a unique set of gene partners. D The unique fluorescent signal combination (across PCR1 and PCR2) for each targeted fusion was validated with cells lines or a synthetic DNA construct.