GSM9380340: Nthy, EZH2 WT, ATAC, rep2; Homo sapiens; ATAC-seq
ATAC-seq was performed as previously described. Briefly, 50,000 cells were harvested and washed with ice-cold PBS. The supernatant was removed, and the cell pellet was resuspended in 50 µL of lysis buffer (10 mM Tris-HCl [pH 7.4], 3 mM MgCl2, 10 mM NaCl, and 0.1% NP-40). Nuclei were collected and resuspended in 50 µL of transposition reaction buffer (10 mM TAPS [pH 8.5], 5 mM MgCl2, and 10% DMF) containing adaptor-loaded Tn5 transposase. The reaction was incubated at 37°C for 30 min in a thermomixer set to 1,000 rpm. To terminate tagmentation, 10 µL of clean-up buffer (900 mM NaCl and 300 mM EDTA [pH 8.0]) was added. Tagmented DNA was purified using the MinElute PCR Purification Kit and quantified via qPCR. Libraries were amplified using the KAPA HiFi HotStart PCR Kit with index primers and purified using SPRIselect DNA Size Selection Reagent. Sequencing was performed on an Illumina NovaSeq X (100 bp paired-end) or NovaSeq X Plus (150 bp paired-end) platform.