title: GSM8490236: WT2, 32kPa, timepoint 7, ATAC-Seq, replicate 2; Homo sapiens; ATAC-seq
library_construction_protocol: Cells were harvested from plates using TrypLE Express Enzyme as described above and stored frozen in liquid nitrogen in Recovery Cell Culture Freezing Medium (Gibco). Once all samples were collected, ATAC-sequencing was performed as previously described. In brief, frozen cells were thawed rapidly in a 37°C water bath and 75,000 cells were aliquoted and washed once with PBS. Nuclei were isolated, lysed, and incubated with Tn5 transposase (Illumina) as described. Transposed DNA was isolated with the MinElute Reaction Cleanup Kit (Qiagen). Libraries were prepared by amplifying DNA for 5 cycles with NEBNext 2x MasterMix (New England Biolabs). After performing qPCR to determine additional cycles needed, libraries were amplified for an additional 6 cycles. Double sided bead purification was performed using AMPure XP beads (Beckman) to remove primers and large (>1000 bp) fragments using standard protocols. Quality and concentration of libraries were determined using a Qubit 4 Fluorometer (Invitrogen) and a 2100 Bioanalyzer Instrument (Agilent). Samples were sequenced on a NovaSeq 6000 (Illumina).