RNA-seq of CD8-positive T cells
Total RNA was extracted from mouse CD8-positive T cells using TRIzol reagent. mRNA was enriched by oligo(dT) magnetic beads and fragmented using fragmentation buffer. First-strand cDNA was synthesized with random hexamer primers, followed by second-strand cDNA synthesis. After end repair and adenylation, Illumina adapters were ligated to the cDNA fragments. The ligated products were size-selected by magnetic beads and amplified by PCR to generate the final sequencing library. Library concentration was quantified by Qubit and fragment size distribution was assessed by Agilent Bioanalyzer.