GSM7025866: B6_NOD_F1_WT_mTEC_GFP_ATAC_rep2; Mus musculus; ATAC-seq
Cells were spun down at 4°C and washed with 1mL PBS supplemented with protease inhibitor (Complete EDTA-free protease inhibitor cocktail, Roche). Then 10uL Tn5 transposase mixture was added to resuspend the cell pellet on ice. [10uL Tn5 transposase mixture: 5uL 2x Tagment DNA Buffer, 0.5uL Tagment DNA Enzyme from the Nextera DNA Library Preparation Kit (Illumina), 4.4uL nuclease-free water, 0.2mg/ml digitonin (G9441, Promega)]. Cells were incubated for 30 min at 37°C with agitation. Transposed DNA fragments were purified using the MinElute Reaction Cleanup Kit (QIAGEN). Two rounds of PCR were performed to generate the library as described in the ImmGen ATAC-seq protocol. After the initial round of PCR with 7 cycles, small DNA fragments were size-selected and purified using the Agencourt AMPure XP beads (Beckman Coulter) followed by a second round of PCR with 10 cycles. Amplified libraries were purified by AMPure XP beads (x1.8 vol.).