GSM8223062: Female, 4 months (T097); Homo sapiens; ATAC-seq
Briefly, fresh human thymus was minced into small pieces and digested in warm dissociation media (2mg/ml Stemxyme, 6.3 U/mL DNaseI, 1.5% BSA in RPMI) for two 30 min intervals. An additional 8 ml of dissociation cocktail was added for a final 30 min digestion. Cells from the third digestion were collected and depleted of CD45+ cells using the Miltenyi LD column prep kit. Cells were then sorted based on live cells and CD45 expression. For some samples fresh thymocytes were enriched for CD25+ cells and and CD8- cells using StemCell Technologies EasySep kits. Cells were recombined in a 3:2 ratio (CD45-:CD45+) or 2:2:1 ratio (CD45-:CD45+:CD25+) prior to library preparation. Nuclei were isolated following the 10X genomics cell line nuclei isolation kit using 0.1X lysis buffer and a 4 minute incubation. All samples were prepared in accordance to the 10X user guidelines (cite user guide) using the ATAC v1 kit.