GSM4794671: CoVi01_4_0_0_5P_TCR; Homo sapiens; RNA-Seq
For combined single-cell RNA-seq and TCR-seq assays (10x Genomics), a maximum of 60,000 virus-reactive memory CD4+ T cells from up to 8 donors were pooled by sorting into low retention 1.5mL collection tubes, containing 500 µL of a1:1 solution of PBS:FBS supplemented with RNAse inhibitor (1:100). Following sorting, ice-cold PBS was added to make up to a volume of 1400 µl. Cells were then centrifuged for 5 minutes (600 g at 4 ˚C) and the supernatant was carefully removed leaving 5 to 10 µl. 25 µl of resuspension buffer (0.22 µm filtered ice-cold PBS supplemented with ultra-pure bovine serum albumin; 0.04 %, Sigma-Aldrich) was added to the tube and the pellet was gently but thoroughly resuspended. Following careful mixing, 33 µl of the cell suspension was transferred to a PCR-tube for processing as per the manufacturer's instructions (10x Genomics). Single-cell RNA-seq using 10x genomics platform was performed using Chromium™ Single Cell 5' v1 Reagent Kits following the manufacturer's protocol Libraries were constructed using Chromium™ Single Cell 5' v1 Reagent Kits following the manufacturer's protocol