title: GSM9094667: ORI_C2C12_ATAC_day4_rep1; Mus musculus; ATAC-seq
library_construction_protocol: The C2C12 myoblasts(ATCC) were cultured in Dulbecco's Modified Eagle's Medium (DMEM)  (Thermo Fisher,10566024) supplemented with 10% fetal bovine serum (Gibco, 10437028) and 1% penicillin-streptomycin (Gibco, 15140122) at 37? in a humidified atmosphere containing 5% CO2 and 95% air ,and were differentiated into myotubes in DMEM containing 2% horse serum (Hyclone, SH30074.03) and 1% penicillin-streptomycin when the cell confluence reaches more than 90%. The differentiation medium should be changed every 24 hours for the first two days of differentiation and every 12 hours after two days. ATAC-seq libraries of mouse testes were prepared as previously described with minor modifications using the High-Sensitivity Open Chromatin Profile Kit 2.0 for Illumina (Novoprotein, N248). Briefly, Approximately 100,000 cells per replicate were processed at three stages: undifferentiated, 48 h, and 96 h differentiation. Cells were PBS-washed and lysed in 50 µl Lysis Buffer (5 min, ice). Nuclei were pelleted (500 g, 5 min, 4°C) after adding 950 µl Wash Buffer. Nuclei were resuspended in 40 µl Tagmentation Mix (37°C, 30 min), then stopped with 10µl Stop Buffer. Fragmented DNA was purified with Novoprotein Tagment DNA Extract Beads (N245) and amplified with NovoNGS Index Kit (Novoprotein, N239). Libraries were purified with Novoprotein DNA Clean Beads (N240), quantified by Qubit, size-verified by Agilent 2100, and sequenced on Illumina NovaSeq 6000.