GSM5463044: ATACseq_H4_D68N_rep1; Homo sapiens; ATAC-seq
MDA-MB-231 cells expressing FLAG-tagged wild-type or mutant histones were seeded in 6-well plates.  For individual samples, 5 x 104 cells were collected, washed, and lysed in 50 μL of cold ATAC Resuspension Buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40, 0.1% Tween-20, and 0.01% digitonin) on ice.  Nuclei were then collected and resuspended in 50 μL of Transposition Mixture (10 mM Tris-HCl pH 7.6, 3 mM MgCl2, 10% dimethylformamide, 100 nM transposase, 0.1% Tween-20, and 0.01% digitonin), and incubated at 37 °C for 30 minutes in a thermomixer at 1,000 RPM. DNA was collected using a Qiaquick PCR purification kit (Qiagen, 28104) according to the manufacturer's instructions.  Eluted DNA was amplified using a KAPA non-hot-start PCR kit and purified using AMPure XP beads. The ATAC-seq libraries were subjected to QC analyses (i.e., the final library yield and the size distribution of the final library DNA fragments) and sequenced using an Illumina NextSeq 500 to an average depth of 57 million unique aligned reads per condition.  Two independent biological replicates were used.