GSM6912132: mESC_HPRT1R_clone1_ATAC_rep1; Mus musculus; ATAC-seq
Cells were grown to medium confluency in 6-well plates. Cells were harvested by washing once with PBS, disso-ciating into single-cell suspending with TrypLE Express (ThermoFisher 12604013) and then neutralizing with mESC media. Cells were counted and 50,000 were taken for tagmentation. Cells were pelleted at 500 rcf for 5 minutes at 4oC, washed with 50 μL cold PBS, resuspended in 50 μL cold ATAC lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630), spun down at 500 rcf for 10 mins at 4oC, resuspended in 50 μL TDE mix. Tagmentation was performed at 37oC on a rotator for 30 mins and DNA was purified using the DNA Clean and Concentrator 5 kit (Zymo Research D4004). PCR was performed as previously described (Buenrostro, J.D., et al., Curr Protoc Mol Biol, 2015) using barcoded primers for 10 total cycles.