GSM8746392: Replicate 1 Day 2 dox; Mus musculus; ATAC-seq
MEFs were seeded onto 0.1% gelatin coated plates and the following day were treated with 1 ng/ml doxycycline for 48 and 96 hours. 50,000 cells of each treated and control cells were centrifuged at 350 x g for 5 minutes at 4°C and washed in cold PBS. The cell pellet was resuspended in 50 μl transposase mixture containing 25 μl 2x TD Buffer, 2.5 μl transposase enzyme, 0.01 % digitonin and 22 μl water. The resuspended cell pellet was incubated at 37°C for 60 minutes at 300 rpm. Subsequently, samples were purified using the MinElute PCR purification kit (Qiagen), according to manufacturer instructions. The elution buffer was pre-warmed to 37 °C and during the final elution step, the buffer was left to incubate on the columns for an additional 5 minutes before the final elution spin step. ATAC-Seq library preparation was performed as described previously (Buenrostro et al., 2015)