GSM8067214
We conducted ATAC-seq based on Omni-ATAC with some modifications. Briefly, 50000 fresh HASMCs were collected and rinsed twice with cold PBS. Cells were spun down at 500 g for 5 min at 4 °C. Cells were resuspended with 50 μL ATAC-seq Lysis Buffer, and incubated on ice for a duration of 3 min. Then we added 1 mL of ATAC-seq Wash Buffer to dilute the lysis reagents. After wash with ATAC-seq Wash Buffer, cells were resuspended with 50 μL Reaction mix (16.5 μL PBS, 0.5 μL 100x cocktail, 0.5 μL 1% Digitonin, 0.5 μL 10% Tween-20, 0.2 μL 20 μM PAT-MEA/B, 10 μL 5x TMgCl-DMF, 21.7 μL ddH2O). Tagmentation was conducted at 37 °C for 30 min at 1000 rpm. To quench Tagmentation, we added 250 μL of DNA Binding Buffer from the DNA Clean and Concentrator-5 Kit and mixed well. Preliminary libraries were purified with the DNA Clean and Concentrator-5 Kit. Follwing Omni-ATAC protocol. After PCR enrichment using 2X KAPA master mix,fragments were purified using the Zymo DNA Clean and Concentrator-5 Kit.