GSM6235178: S9CC47_R5_1p6en8; Homo sapiens; ATAC-seq
ATAC-seq was performed as described previously (Corces et al., 2017). Briefly, CNCCs were incubated with BCh media containing 200U/mL DNaseI (Worthington, LS002007) for 30 min AND harvested using accutase. Viable cells were counted using Countess Automated Cell Counter (Invitrogen), 50,000 viable cells were pelleted at 500 RCF for 5 min at 4°C and resuspended in ATAC-resuspension buffer containing 0.1% NP40, 0.1% Tween20, and 0.01% Digitonin and incubated on ice for 3 minutes. Following wash-out with cold ATAC-Resuspension Buffer (RSB, 10 mM Tris-HCl pH 7.4, 10mM NaCl, 3mM MgCl2 in sterile water) containing 0.1% Tween20, cells were pelleted and resuspended in 50 μL transposition mix (25 μL 2x TD buffer, 2.5 μL transposase (100nM final), 16.5 μL PBS, 0.5 μL 1% digitonin, 0.5 μL 10% Tween20, 5 μL H2O) and incubated for 30 minutes at 37°C with shaking. The reaction was purified using the Zymo DNA Clean & Concentrator kit Purified, transposed DNA was PCR-amplified with NEBNext master mix and primers as defined in Corces et al. Libraries were purified by two rounds of double-sided size selection with AMPure XP beads (Beckman Coulter, A63881), with the initial round of 0.5x sample volume of beads followed by a second round with 1.3x initial volume of beads. Library size distributions were confirmed by separation on a PAGE gel and staining with SYBRGold and pooled based on quantifications from Qubit dsDNA High Sensitivity Kit.