A reference model for circulating hematopoietic stem cells with applications to diagnostics

We performed deep targeted somatic mutation analysis to identify cases of clonal hematopoiesis (CH) associated with pre-leukemic mutations. For the healthy cohort, we used our CH panel V3, containing 705 probes, covering leukemia-related Single Nucleotide Variants (SNVs) and Indels in 47 genes, complemented by two amplicon sequencing reactions to cover GC-rich regions in SRSF2 and ASXL1. For the cytopenic cohort, we used our CH panel V4 (described in detail in Biezuner, T. et al., NAR Genom Bioinform, 2022). Both panels were designed to ensure capture uniformity and specificity. Each DNA sample was sequenced twice with a minimum depth of 1,000,000 paired-end reads on an Illumina Novaseq 6000 machine.