GSM5741887: MEC1_crispred_ALT_ATACseq_rep2; Homo sapiens; ATAC-seq
50,000 cells were washed with PBS, resuspended in 50 μl RSB buffer (10 mM Tris-HCL pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP40, 0.1% Tween20) and incubated on ice for 3 min. Cells were spun down, supernatant was discarded, cells were resuspended in 1 ml RBS buffer without NP40, inverted three times and spun down. Supernatant was discarded, cells were resuspended in 25 μl 2X Tagmentation buffer (20 mM Tris-HCl pH 7.6, 10 mM MgCl2, 20% Dimethyl Formamide), 0.5 μl 10% Tween20, 16.5 μl PBS, 1 μl of homemade 24.5 μM Tn5 and 7 μl H2O and incubated 30 min at 37ºC rotating at 1000 rpm. DNA was then purified with a DNA Clean & Concentrator-5 (Zymo Research). Tagmented and amplified 11 cycles.