GSM4909460: CNCC_ATAC_100_bio3; Homo sapiens; ATAC-seq
ATAC-seq libraries were generated from whole cells as previously described (Calviello et al., 2019). Briefly, cells were dissociated with StemPro Accutase and 100,000 viable cells (as determined using the Muse Cell Analyzer (Merck) 'Count and Viability' assay) were washed in ice cold PBS. Cells were pelleted at 500 xg for 5 min at 4 °C and resuspended in transposition reaction mix (25 µL 2 x TD reaction buffer (Illumina), 2.5 µL Tn5 transposase (Illumina) and 22.5 µL sterile water) and incubated at 37 °C in a thermoshaker at 1,400 rpm for 1 hour. The transposition reaction was immediately purified in 10 µL elution buffer using a MinElute reaction clean-up kit (Qiagen). Transposed DNA fragments were then initially amplified using 2.5 µL each of Nextera Index i7 and Nextera Index i5 (Nextera index primer kit, Illumina) with 25 µL NEB Next High Fidelity 2 x PCR master mix (NEB) under the following conditions: 72 ° for 5 min, 98 °C for 30 sec followed by five cycles of 98 °C for 10 sec, 63 °C for 30 sec and 72 °C for 1 min. To reduce GC and size biases, the total PCR cycle was determined as previously described