Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: CD8+ T cells were isolated from LCMV-infected mice by CD8+ MACS negative selection performed on single-cell suspensions of splenocytes, followed by FACS. CD8+ T cells were isolated from B16-ova tumors by dissecting tumors from the surrounding fascia, mechanically mincing and treating with collagenase P and DNAse I for 10 minutes at 37C. Tumor-infiltrating leukocytes were enriched by CD8+ MACS positive selection (Miltenyi) and FACS isolation. In both contexts, cells were sorted directly into RLT buffer + 1% BME in 96 well plates. Library construction was performed according to a modified Smart-Seq2 protocol as described in Yates et al., PNAS, 2018.