GSM9126517: eEE biol_rep 1 and 2; Mus musculus; ATAC-seq
Epithelia (esophageal control, eEE; and esophageal-derived skin, eSKIN) were harvested 10 days post-culture, at which point the epithelia were carefully peeled from their underlying stroma following a 50mM EDTA incubation for 15min at 37°C. The peeled epithelia was rinsed with PBS, and incubated with 0.5mg/ml Dispase (Sigma) for 5min. EDTA was then added to the samples at a final concentration of 5mM, and the suspension diluted 1/5 by adding PBS + 0.04% BSA buffer. A cell suspension was obtained by filtering through a 30um cell strainer, centrifuging at 300g for 5min at 4 degrees and resuspending in PBA + 0.04% BSA. After assessing viability and cell number, nuclei were isolated following 10X CG000169 protocol. Libraries were prepared following manufacturer's instructions for Chromium Next GEM Single Cell ATAC Kit v2