GSM6895895: f-Hep-2 ATAC-seq; Homo sapiens; ATAC-seq
f-Heps, mid-Heps, and a-Heps were dissociated with Accutase, and collected in HCM medium. Frozen PHHs purchased from Celsis IVT (Lot: ONO) were recovered according to the manufacturer's instructions. ATAC libraries were prepared using Omni-ATAC protocol as previously described, and TruePrep® DNA Library Prep Kit V2 for Illumina (Vazyme, China) was employed. Briefly, 50,000 cells were washed with cold PBS once and lysed with 50 μl cold ATAC-Resuspension Buffer (RSB) (10 mM pH 7.4 Tris-HCl, 10 mM NaCl, and 3 mM MgCl) containing 0.1% NP40 and 0.1% Tween-20. After incubation on ice for 3 minutes, 1 ml of cold ATAC-RSB containing 0.1% Tween-20 (without NP40) was added to the cell lysate and mixed by inverting 3 times. The nuclei were collected by centrifuging at 500 g for 10 minutes at 4 °C. For the transposition reaction, nuclei were incubated with Tn5 Transposition mix (10 μl 5 × TTBL buffer, 3 μl TTE Mix V50 transposase, 16.5 μl PBS, 0.5 μl 10% Tween-20 and 20 μl H2O) at 37 °C for 30  minutes. Next, the stop buffer was directly added to end the reaction, and  PCR was performed to amplify the library for 12 cycles. Then the libraries were purified with 1.2 × AMPure beads (Beckman), and sequenced using an Illumina Novaseq 6000.