title: GSM6249027: 5ABDCB, 0, Exposed, Exposed [ATAC-Seq]; Homo sapiens; ATAC-seq
library_construction_protocol: Nuclei were extracted from frozen PBMCs. Briefly, 100K cells were spun down at 300 x g for 5 minutes at 4oC. The supernatant was removed, and cells were mixed with 100 uL of lysis buffer (10mM NaCl, 3mM MgCl2, 10mM Tris-HCl pH7.4, 0.1% Tween-20, 0.1% NonidetTM P40) and lysed on ice for 4 minutes. Wash buffer (1 mL; 10mM NaCl, 3mM MgCl2, 10mM Tris-HCl pH7.4, 0.1% Tween20) was added before centrifuging at 500 x g for 5 minutes at 4oC. Transposition mix (25 μL 2× TD buffer, 2.5 μL transposase (Tn5, 100 nM final), 22.5 μL water) was added to the nuclear pellets, incubated at 37 °C for 30 minutes, and samples purified using the Qiagen MinElute PCR Purification Kit. DNA fragments were PCR amplified for a total of 10-11 cycles and resulting libraries purified using the Qiagen MinElute PCR Purification Kit.