title: ATAC-seq ofTcf-1 deficient CD4+CD8+ thymocytes
design: 1x105 CD4+CD8+ T-cells sorted from CD4-Cre/Tcf-1fl/- mice were used for ATAC-seq. Cells were centrifuged at 500g at 4C for 5 minutes, washed with 1X PBS, and centrifuged again. Cells were resuspended in lysis buffer (10mM Tris-HCl pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% IGEPAL CA-630) and immediately centrifuged at 500g at 4C for 10 minutes. Pellets were resuspended in transposition reaction buffer (25l 2x Tagment Buffer (FC-121-1030, Illumina), 2.5l Tagment DNA Enzyme, 22.5l nuclease free H2O) for 30 minutes at 37C. DNA was purified with a Qiagen MinElute Kit and amplified with Nextera PCR Primers (Illumina Nextera Index Kit) and NEBNext PCR Master Mix (M0541, New England BioLabs) for 11 cycles. Amplified DNA was purified with a Qiagen PCR cleanup kit