GSM9790644: OSC19 cells (oral quaous cell carcinoma cells), treated with Tryp metabolites, rep 2; Homo sapiens; ATAC-seq
Cells were incubated with DNase enzyme (20,000 IU/ml) in 1x HBSS (supplemented with 250 mM MgCl2 and 50 mM CaCl2) for 30 min in 37°C incubator. Cells were washed with 1x PBS, detached using Trypsin-EDTA solution (Sigma), neutralized with FBS-containing media, resuspended in cold 1xPBS, counted, and frozen 150,000 cells per replicate in freezing media (50% FBS, 40% DMEM/F12, 10% DMSO) at -80°C. Triplicate samples per treatment group were submitted to ActiveMotif, Inc (Carlsbad, CA) for ATAC-seq analysis. Frozen cells were thawed, diluted in RPMI medium, pelleted, washed with cold 1x PBS, and subjected to Tn5-mediated tagmentation as previously described using the Active Motif ATAC-Seq Kit. Tagmented DNA was purified using the MinElute PCR Purification Kit (Qiagen), amplified for 10 PCR cycles, cleaned using Agencourt AMPure SPRI beads (Beckman Coulter), libraries quantified using Qubit (Invitrogen) and TapeStation (Agilent), and sequenced as PE50 reads on an Illumina NovaSeq X Plus platform.