GSM8219606: SK-L012-HEK293T-ATAC_Kethoxal-2; Homo sapiens; ATAC-seq
ATAC-seq experiments were carried out following the Omni-ATAC protocol. Briefly, ~50,000 cells were centrifuged at 5,00 g, then resuspended in 500 UL 1x PBS and centrifuged again. Cells were then resuspended in 50 UL ATAC-RSB-Lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630, 0.1% Tween-20, 0.01% Digitonin) and incubated on ice for 3 minutes. Subsequently 1 mL ATAC-RSB-Wash buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.01% Digitonin) were added, the tubes were inverted several times, and nuclei were centrifuged at 500 g for 5 min at 4C. Transposition was carried out by resuspending nuclei in a mix of 25 uL 2X TD buffer (20 mM Tris-HCl pH 7.6, 10 mM MgCl2, 20% Dimethyl Formamide), 2.5 uL Tn5 transposase (custom produced) and 22.5 uL nuclease-free H2O, and incubating at 37C for 30 min in a Thermomixer at 1000 RPM. Transposed DNA was isolated using the MinElute PCR Purification Kit (Qiagen Cat# 28004/28006), and PCR amplified (72C for 3 minutes, 98C for 30 seconds, 10 cycles of 98C for 10 seconds, 63C for 30 seconds, 72C for 30 seconds). Libraries were purified using the MinElute kit, then sequenced on a Illumina NextSeq 550 as 2x38mers.