title: GSM6261559: PD1 treated GP33+ CD8 T cells from LCMV chronically infected mice; Mus musculus; RNA-Seq
library_construction_protocol: Cell sorting was performed byFACS Aria II (BD Biosciences).For scRNA-seq (10X Genomics), LCMV chronically infectedmice (> day 40 p.i.) were untreated or treated with various therapeutic modalities for 2 weeks, and DbGP33+CD8+T cells in spleens were sorted from pooled spleens. Naïve (CD44lo) CD8+T cells were sorted from pooled spleens from uninfected mice (n=2). Single cell RNAseq libraries were generated using the Chromium Single Cell 5'1007Library & Gel Bead Kit (10x Genomics) according to the manufacturer's protocol. Briefly, DbGP33+CD8+T cellsor naïve CD44loCD8+T cells were sorted and captured into the Gel Beads-in-emulsion (GEMs). After reverse transcription, the GEMs were disrupted and the barcoded cDNA was isolated, pooled, and amplified by PCR (13cycles). The amplified cDNA was fragmented, and subjected to end repair and A-tailing followed by a sample index PCR (16 cycles). The purified libraries were sequenced to a depth of 50,000 reads/cell on a 1013HiSeq3000 (Illumina) with 26 cycles for read 1, 8 cycles for index 1 (i7), and 91 cycles for read 2.