GSM8155570: BRM014, BT549 cells, Biorep2; Homo sapiens; ATAC-seq
50,000 cells were lysed in ATAC-RSB (resuspension buffer) (10mM Tris-HCL pH 7.4, 10mM NaCl, 3mM MgCl2, 0.1% Tween-20) supplemented with 0.1% IgePal-CA630 and 0.01% Digitonin before being washed three times in cold ATAC-RSB buffer. Nuclei were pelleted and resuspended in 50μL transposition mixture (17.75μL PBS, 5uL Nuclease Free Water, 25uL TD 2x reaction buffer, 0.5uL 1% digitonin, 0.5uL 10% Tween-20, 1.25μL TDE1 (Illumina, 20034197)) and incubated at 37 degrees in a thermomixer at 1000RPM for 30min. DNA was cleaned using Qiagen PCR Purification kit before PCR amplification for 10 cycles using Illumina index primers, 25μL Q5 High Fidelity 2x master mix (New England BioLabs, M0492) in a final volume of 50μL. Samples were size selected and purified using Ampure XP beads (Bechman Coulter, A63881). Next-generation sequencing was performed using Illumina Novaseq 6000 to get ~25M 50bp paired-end reads per sample