GSM5831326
Cells were processed following the 10X Genomics scATAC-seq protocol. Briefly, ~50,000 sorted mTECs were spun down and examined for cell number and viability. Nuclei were isolated in 50ul ice-cold lysis buffer (10mM Tris-HCl [pH 7.4], 10mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.1% NP-40, 0.01% digitonin, 1% bovine serum albumin) for 5 minutes (first replicate) or 1 minute (second replicate). Nuclei were washed and resuspended in nuclei buffer to target 5,000 recovered nuclei per replicate. Nuclei were transposed at 37°C for 60 minutes, loaded into gel beads-in-emulsion, and incubated for linear amplification. Barcoded single nucleus fragments were purified and amplified by polymerase chain reaction (PCR; 12 cycles).