GSM7316474: CD8Tm 5' Gene expression; Homo sapiens; RNA-Seq
CD8 memory and naive T cells were freshly isolated from Peripheral blood mononuclear cells by positive selection with CD8 MicroBeads (Miltenyi Biotec) with autoMACS Pro Separator and by antibodies cell sorting with CD8+ CD45RA- lymphocytes with BD FACSAria III. Fresh PBMCs were labelled with CD8 MicroBeads at RT for 15min. PBMCs were washed and resuspended with MACS buffer (PBS, 1% v/v FCS, 2 mM EDTA). Beads labelled PBMCs proceeded with autoMACS program positive selection in sensitive mode. MACS-purified CD8 cells were stained with fluorochrome-conjugated antibodies, washed and resuspended in MACS buffer. CD8 T cells were sorted using BD FACSAria III (BD Biosciences) according to the presence of CD8 and the absence of CD45RA antibodies. The sorted CD8 T cell subsets were washed in FCS-contained PBS (10% v/v FCS). Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index)