WGS of Epithelioid-like Tumor 2
Tumor resections were gently minced using scissors into 4 mm pieces and enzymatically dissociated using a cocktail of 1.5 U/mL Collagenase D (Roche 11 088 882 001) and 2.4 U/mL Dispase II (Roche 04 942 078 001) for 60 minutes. The resultant cell suspension was passed through a 40um filter and centrifuged at 500g for 5 mins. Cells were then incubated for 30 mins on ice with the following lineage antibody cocktail [anti-CD45-Alx647 (Ablab 67-0047-01, 1:400), anti-CD31-APC (BD Biosciences 551262, 1:400), anti-CD11b-Alx647 (Ablab 67-0055-01, 1:500), anti-F4/80-Alx647 (Ablab 67-0035-05, 1:500), anti-CD117-APC (eBioscience 17-1172-82, 1:500) and anti-Ter119-647 (Ablab 67-0031-01, 1:200) dissolved in FACS buffer (PBS with 2% FBS and 2mM EDTA)] before fluorescence-activated cell sorting (FACS) against lineage markers to reduce the number of contaminating cell types and for tdTomato positive cells on an Influx (BD) sorter. Samples were prepared using the DNA PCR-Free Library Tagmentation Prep kit (Illumina). The libraries were sequenced on the HiSeq-X platform.