GSM8718624: hPSC_ATAC_3; Homo sapiens; ATAC-seq
Nuceli were extracted using a modified version of the OmniATAC-seq protocol. 50,000 live, freshly isolated cells were washed with 50 µl cold 1X PBS and pelleted. 50 µl of cold lysis buffer (0.1% v/v NP-40, 0.1% v/v Tween-20, 0.01% v/v Digitonin in resuspension buffer, comprised of 10 mM Tris-HCL, 10 mM NaCl, and 3 mM MgCl2) was added to each sample and incubated for 3 minutes on ice. 1 mL of wash buffer (resuspension buffer with 0.1% v/v Tween-20) was added to each sample before centrifugation at 500xg for 10 minutes at 4C. The supernatant was discarded, and each nuclei pellet was resuspended in 50 µl of transposition buffer (25 µl 2X TD Buffer, 16.5 µl 1X PBS, 0.5 µl 10% Tween-20, 0.5 µl 1% Digitonin, 2.5 µl Tn5 Transposase, 5 µl nuclease-free water). The mixture was incubated for 30 minutes at 37°C on a thermomixer set to 1000 rpm. Transposed DNA was isolated using the Qiagen MinElute Reaction Cleanup Kit, as per manufacturer's instructions. Libraries were generated using Illumina Nextera DNA Unique Dual Indexes and NEBNext High-Fidelity 2X PCR Master Mix. Libraries were double-side purified using AMPure XP beads, and quality was assessed using Agilent Bioanalyzer (High Sensitivity DNA kit). Sequencing was conducted on a NovaSeq 6000.