GSM8706160: BT37_Corin_ATAC-seq_1; Homo sapiens; ATAC-seq
BT37 and CHLA06 cells were seeded 105,000 cells/cm2 and treated with 1 µM Corin and DMSO for 24 h. 100,000 viable cells were aliquoted, and samples were prepared using ATAC-seq kit (Active Motif, 53150). Cells were pelleted at 500 x g for 5 min at 4 °C, washed with PBS and resuspended in ATAC Lysis Buffer followed by centrifugation at 500 x g 10 min at 4 °C. Pelleted nuclei were resuspended in Tagmentation Master Mix (Tagmentation Buffer, PBS, 0.01% Digitonin, 1% Tween 20, H2O and Assembled Transposomes) and incubated at 37 °C for 30 minutes in a thermomixer at 800 rpm. Then, DNA Purification Binding Buffer was added to the samples along with 3 M sodium acetate as required to reach the appropriate pH for binding. Samples were transferred to DNA purification columns and washed with Wash Buffer and eluted with DNA Purification Elution Buffer. Centrifugation of samples were done at 17000 x g for 1 min. Collected samples were amplified using unique combination of index primers with the following cycling conditions: 72 °C for 5 min, 98 °C for 30s, 10 cycles of 98 °C for 10s, 63 °C for 30s and 72 °C for 1min, hold at 4°C. SPRI beads were added to each sample and vortexed and incubated for 5 min for binding and magnetically separated and washed with 80% ethanol solution twice while remaining attached. Following ethanol evaporation, beads were vortexed in DNA Purification Elution Buffer and incubated for 5 min at RT followed by collection of eluted samples. Library was prepared using ATAC-seq Kit (53150). Sequencing was conducted by Azenta Life Sciences.