GSM7898166: tt2iLIFGo, dm-aKG, rep3; Homo sapiens; ATAC-seq
For ATAC-seq previous protocol was used (Grandi, Modi et al. 2022). Cells were trypsinized and resuspended in N2B27. After washing in PBS 50'000 cells were lysed for 3min on ice in lysis buffer (0.1% NP40; 0.1% Tween-20; 0.01% Digitonin; 10 mM Tris-HCl pH 7.4; 10 mM NaCl, 3 mM MgCl). Lysis was stopped with ATAC-wash buffer (0.1% Tween-20; 10 mM Tris-HCl pH 7.4; 10 mM NaCl, 3 mM MgCl). After pelleting the cells were resuspended in transposition mix [1x TD buffer (Illumina, cat. no. 20034197), 0.01% Digitonin; 0.1% Tween-20 in PBS; TDE1 TD enzyme (Illumina, cat. no. 20034197)] and incubated for 30min at 37°C in a thermomixer set on 1000 rpm. Transposition was terminated with DNA Binding Buffer DNA Clean and Concentrator-5 kit (Zymo Research, cat. no. D4014) and DNA was purified. Libraries were prepared using barcoding adapters and the NebNext High-Fidelity 2x Master Mix (NEB cat. No. M0541S). Cycling conditions used were: 72°C 5min; 98°C 30sec; 11x(98°C 30sec; 63°C 30sec); 72°C 1min. Libraries were purified twice using DNA Clean and Concentrator kit.