GSM9626416: 193-MDCK-wt-B; Canis lupus familiaris; ATAC-seq
50,000 cells was collected and centrifuged at 500 x g for 5 min, 4°C. Cells were washed once with 50 µL of cold 1xPBS buffer and centrifuged again. Supernatant was removed, and cells resuspended in 50 µL of cold lysis buffer followed by centrifugation with the same settings. Supernatant was removed. Libraries were produced as described in Buenrostro, J., Giresi, P., Zaba, L. et al. Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position. Nat Methods 10, 1213–1218 (2013). https://doi.org/10.1038/nmeth.2688. Briefly, a transposition mix (25 μL 2× TD buffer, 2.5 μL transposase (Tn5, 100 nM final), 22.5 μL water) was added to the nuclear pellet. Reaction was incubated at 37 °C for 45 minutes and amplified using PCR. Samples were purified using Qiagen MinElute PCR Purification Kit and again using Agencourt AMPure XP magnetic beads.