Targeted DNA sequencing WM989 cell line
We sequenced WM9-A6 (main WM9 subclone), intermediate subclone A6-B1, and two resistant subclones, A6-B1-A2 and A6-B1-A3. We obtained resistant subclones by isolating cell clusters and expanding in the presence of drug. For each cell line, 500 ng of genomic DNA was sheared randomly into 200 bp fragments with the CovarisTM S200 UltraSonicator (Covaris¨). ĘSheared DNA was A-tailed and ligated with adaptor-embedded indexes using the NEBNext¨ UltraTM DNA Library Prep Kit for Illumina¨ (New England BioLabs, Inc.). ĘDNA quality, fragment size, and concentration of library preps were measured using AgilentŐs DNA 1000 chips in conjunction with the 2100 Bioanalyzer (Agilent Technologies). Samples were equimolarly pooled and sequenced with a 2.318Mbp SureSelectXT Custom Target Enrichment Kit (Agilent Technologies) targeting 119 genes (Extended Data Fig. 1). Paired-end sequencing (2x125bp) was carried out on the HiSeqTM 2500 sequencing system (Illumina) at the next Generation Sequencing Core Facility. Mean target coverage of 346X was achieved.