title: GSM2966340: True naive CD8 cells timestamped at day 28, rep 1; Mus musculus; RNA-Seq
library_construction_protocol: CD8+ cells were magnetically enriched with CD8 microbeads and sorted to > 95% purity with a FACS Aria III on CD4- CD8+ TdTomato+ CD44 hi (virtual memory) or lo (true naïve) populations. Total RNA was isolated with Trizol, with an extra chloroform extraction to remove residual phenol and addition of glyco-blue as a carrier to promote RNA precipitation. Directional RNA-seq libraries were prepared from total RNA isolated from 10,000 cells (~10-20ng) using the NEBNext Directional Ultra II RNA Library Prep Kit for Illumina (New England Biolabs), with initial polyA+ isolation.