GSM6209212: 27-CLP-6; Homo sapiens; ATAC-seq
At endpoint, cells were harvested for ATAC-seq. ATAC-seq was performed as previously described (Corces et al., 2017) with modifications. Specifically, 50,000 cells were resuspended in 500 μL resuspension buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, and 3 mM MgCl2 in water). Cells were centrifuged at 50xg for 10 min in a pre-chilled (4°C) fixed-angle centrifuge. Lysis and transposition were performed simultaneously by resuspending cells in 25 μL tagmentation mix (12.5 μL 2xTD buffer [Illumina, Cat. No. 15027866], 1.25 μL Tn5 enzyme [Illumina, Cat. No. 15027865], 9.25 μL PBS, 0.5 μL 1% digitonin, 0.5 μL Tween-20, 0.5 μL NP40, and 0.5 μL 100x protease inhibitors), and incubating at 37°C for 30 min in a thermomixer with shaking at 1,000 rpm. Tn5 transposase-tagged DNA was amplified by PCR and purified using AMPure XP magnetic beads. ATAC-seq reads were sequenced 2x150 bp using a NovaSeq 6000. Sequencing of ATAC-seq libraries was performed by the University of Michigan Advanced Genomics Core.