design: ATAC-seq of CD44+CD8+ T cells isolated from spleen on 5 post rechallenge in a heterotypic influenza rechallenge model
library_construction_protocol: CD3+44+8+ T cells were isolated from splenocytes on day 5 after restimulation with a heterotypic strain of influenza, having received primary infection 30 days previously along with either sodium valproate (0.1%w/v) in drinking water or vehicle (1 in 10,000 DMSO). Following CD8+ T cell isolation 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.