GSM7890032: Resistant_304-rep2; Homo sapiens; ATAC-seq
Cell lines were lysed on-plate for RNA isolation using the Qiagen RNeasy Plus Mini kit (Cat. No. 74034), and RNA concentration was determined using a NanoDrop 8000 (Thermo Fisher Scientific).  RNA integrity was confirmed using a “Bioanalyzer 2100 (Agilent Technologies). Briefly, cryo-preserved cells were thawed, washed with 1X PBS and counted. About 100,00 cells were then lysed for 5 min on ice in 100 µl lysis buffer (10 mM Tris (pH 7.5), 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40, 0.1% Tween- 20 and 0.01% digitonin). 1ml wash buffer (10 mM Tris (pH 7.5), 10 mM NaCl, 3 mM MgCl2 and 0.1% Tween-20) was then added and the pellets were spun down at 500g for 10 min at 4°C. The supernatant was discarded and the pellet was resuspended in 50 µl tagmentation buffer (25 µl 2X TD buffer, 16.5 µl PBS, 0.5 µl 10% Tween-20, 0.5 µl 1% digitonin, 2.5 µl Tn5 transposase and 5 µl H2O). The tubes were incubated at 37 °C for 30 min, followed by DNA isolation using the Qiagen MinElute Cleanup Kit (catalog number 28206). The DNA was then amplified using NEBNext 2X PCR Master mix (catalog number M0541L) and analyzed by TapeStation (Agilent) before submission for sequencing.