2.9.1. Droplet digital PCR assay design

ddPCR assays for SNVs/indels, with probes labelled with 5’-FAM™ or 5’-HEX™ fluorophores and 3’-Iowa Black^® Fluorescent Quencher (FQ), were obtained from Bio-Rad (Bio-Rad, CA, US) ( Table S6 ). Custom ddPCR assays for SVs, copy-number reference (CNR) assay Albumin and 1 SNV (IRF4 p.L24V) were designed using Primer3Plus web interface (Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology), according to Bio-Rad guidelines ( Table S7 ). Dark probes i.e., probes labelled with only 3’-Iowa Black^® FQ, were designed to target the wild-type allele for some Bio-Rad ddPCR assays using the MIQE context sequence of the corresponding Bio-Rad assay ( Table S6 ). Custom designed primers and HPLC purified probes, with 5’-FAM™ or 5’-HEX™ fluorophores and 3’-Iowa Black^® Fluorescent Quencher, and HPLC purified, were ordered from IDT (Integrated DNA Technologies) ( Table S7 ). Locked nucleic acid (LNA) bearing Affinity Plus^® probe was ordered for IRF4 p.L24V ( Table S7 ).