design: ATAC-seq of isolated primary human CD8+ T cells treated with KDACi with polyclonal stimulation for 6 days
library_construction_protocol: CD8+ T cells were isolated by positive magnetic selection from leucocyte cones from healthy blood donors before being stimulated with anti-CD2/3/28 microbeads in complete RPMI medium with non-limiting IL2 (10ng/ml) in the presence of either sodium valproate (175uM) or vehicle (1 in 10,000 DMSO) Following CD8+ T cell culture as above, 50,000 cells were lysed (1M Tris-HCl) and the nuclear pellet transposed using the Tn5 transposase (TDE1, Illumina) at 37C for 30min before DNA isolation (Qiagen minElute) and PCR amplification (NEBnext) guided by qPCR (KAPA-SYBR) estimation of reporter signal v cycle number and PCR cycle length tailored to 1/3 maximal amplification Libraries were purified (AMPure XP) with quality assessed using an Agilent BioAnalyser 2100 and RNA quantification performed using a NanoDrop ND-1000 spectrophotometer.