GSM5058856: OD12_Positive; Homo sapiens; RNA-Seq
Fresh peripheral blood T cells were enriched for CD4+ T cells using the Easysep human CD4+ enrichment kit. Enriched CD4+ T cells were then stained with CMV Class II tetramer, and subsequently surface antibodies for 15 minutes at 4oC. Using the MoFlo™ sorter, CD3+, CD4+, tetramer+ cells were sorted into 2 populations; PD1+  and PD1-. Sorting was performed within 4 hours of blood sampling.  After sorting, cells were immediately pelletted,  3500g for 10 minutes, and RLT plus lysis buffer added and vortexed for 1 minute. RNA extraction was then performed according to manufacturer's instructions. (RNEasy Plus micro kit, Qiagen). RNA libraries were prepared using a TruSeq Stranded Total RNA Library preparation kit RNA-Seq - Illumina HiSeq2000 platform using TruSeq version 3 chemistry, over 100 cycles (Oxford Gene Technologies).