title: NextSeq 550 paired end sequencing; RNAseq analysis of CD4 and CD8 T cells in response to vaccinia virus infection
design: RNAseq analysis of CD4 and CD8 T cells in response to vaccinia virus infection
library_construction_protocol: CD8+CD44highCD62L-GFPEgr2high cells were isolated from Egr2 Kin mice by FACS GFP-Egr2 knockin (Egr2 Kin) and hCD2-Cre Egr2loxP/loxP Egr3-/- (Egr2/3 DKO) mice were infected intranasally with 2 x 105 PFU of vaccinia virus in 10ul of physiological saline. CD4 and CD8 T cells were isolated 7 days after infection RNA was isolated and purified using TRIzol reagent (Life Technologies) according to the manufacturer's instructions RNA from T cells was processed for directional mRNA-Seq library construction using the TruSeq Stranded mRNA Library Prep Kit (Illumina) according to the manufacturer's protocol