GSM8267184: CTRL2_IFN_REP2 [ATAC-Seq]; Homo sapiens; ATAC-seq
ECs were dissociated by trypsin, and 50,000 cells were collected in PBS. A 50 µl of reaction mix containing Transposase enzyme, buffer (Illumina Tagment DNA TDE1 enzyme and buffer kit), and Digitonin (2%) (Promega) was added to the cell pellet as recommended (Corces et al., 2016). The reaction was incubated at 37 oC for 30 min with agitation at 300 rpm, before purifying with a Qiagen MinElute PCR purification kit. The transposed DNA was stored at -20 oC to wait until all samples were ready for PCR amplification. The transposed DNA fragments were amplified for 4-6 cycles (NEBNext High-Fidelity 2X PCR Master mix), before being cleaned again with a Qiagen MinElute PCR purification kit. DNA fragments (100-1000 bp) were selected from the library by bead purifications (Agencourt AMPure XP beads, Fisher Scientific). The quantity and quality of the ATACseq libraries were calculated by Bioanalyzer (Agilent TapeStation), HS D1000. The libraries were pooled together and sequenced by BGI on the DNBSEQ platform with dual index.