GSM5020850: 24hr_K3_Uninfected_Rep2; Homo sapiens; ATAC-seq
Cells were collected and resuspended in 50 μl of a transposase mix that included 25 μl of 2x Nextera Tagment DNA buffer (Illumina), 2.5 μl Nextera Tn5 transposase (Illumina), 0.5 μl digitonin 5% (Invitrogen), and 22μl of H2O. The reaction was carried at 37°C for 30 min. DNA was purified with Qiagen MinElute PCR purification kit and eluted in 10μl of buffer EB. DNA libraries were amplified by using NEBNext High-Fidelity (New England Biolabs) and barcoded primers. The number of amplification cycles were calculated by qPCR and kept under 11 to prevent saturation. Libraries were purified with Qiagen MinElute PCR purification kit and sized selected with AMPure XB beads (Beckman Coulter) . Library quality control was assessed by Bioanalyzer High Sensitivity DNA Analysis kit (Agilent) and the libraries were sequenced with Illumina NextSeq technology (75bp single-end reads, 100 million reads/sample).