GSM1526681: naive CD8+ T-cells; Homo sapiens; RNA-Seq
Venous peripheral blood (60 ml) from a healthy donor was collected in a lithium-heparin BD Vacutainer tube (BD, Franklin Lakes, NJ, USA). Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll Paque Plus (GE Healthcare Life Sciences, Uppsala, Sweden) gradient centrifugation and subjected to staining for FACS analysis. The red blood cells in the pellet were lysed with monochloride solution (155 mM NH4Cl, 10 mM KHCO3, 0.1 mM Na2.EDTA.2H2O, pH 7.4) to yield the granulocyte fraction.The PBMCs were incubated with antibodies for 45 minutes at 4°C and sorted in six different populations on the MoFloTM XDP flow cytometer (Beckman Coulter, Brea, CA, USA). RNA was extracted from all seven immune cell types using the MirVana RNA isolation kit (Ambion, Life Technologies, Carlsbad, CA, USA) according to the manufacturer’s instructions. sequencing libraries were prepared from 1 µg of total RNA using the TruSeq RNA kit (Illumina) according to the manufacturer’s instructions.
GEO Accession
GSM1526681