GSM8603680: SBC5_SH1; Homo sapiens; ATAC-seq
For each submitted sample, 60 000 viable cells were pelleted at 500 x g for 5 minutes at 4oC, resuspended in 50 µL of fully supplemented ATAC-seq resuspension buffer (RSB) containing 0.1% NP-40, 0.1% Tween-20, and 0.01% digitonin, and incubated on ice for 3 minutes. Following lysis, 1 mL of ATAC-seq RSB containing 0.1% Tween-20 (without NP-40 or digitonin) was added and the tubes were inverted to mix. Nuclei were then centrifuged at 500 x g for 10 minutes at 4oC, followed by careful aspiration of all supernatant and resuspension of pellet with 50 µL of transposition master mix (25 µL 2x TD buffer, 2.5 µL transposase, 16.5 µL PBS, 0.5 µL 1% digitonin, 0.5 µL 10% Tween-20, and 5 µL water). Transposition reactions were incubated at 37oC for 30 minutes in a thermomixer with shaking at 1000 rpm. Reactions were cleaned up with MinElute columns (Qiagen, catalog # 28204), followed by library amplification, where the appropriate number of PCR cycles was determined by qPCR. PCR product purification and size selection were achieved using AMPure XP Beads (Beckman Coulter, catalog # A63882).