Tom+ LCMV-specific TRM cells day >30 - Liver - rep 3
In brief, total RNA was extracted using the standard TRIzol (Invitrogen) protocol and used for library preparation and sequencing. mRNA was processed as described by Hashimshony et al. Cell Rep 2012 and Simmini et al. Nat Comm 2014, following an adapted version of the single-cell mRNA seq protocol of CEL-Seq. Samples were barcoded with CEL-seq primers during a reverse transcription and pooled after second strand synthesis. The resulting cDNA was amplified with an overnight in vitro transcription reaction. From this amplified RNA, sequencing libraries were prepared with Illumina Truseq small RNA primers. Paired-end sequencing was performed on the Illumina Nextseq500 platform.