GSM9705102: TCR-seq, T-cells, VDJ; Homo sapiens; OTHER
Cells were washed three times through sedimentation at 390 g and resuspended in PBS+0.5% BSA before sorting single, live, CD8+ and tetramer positive cells on a BD FACS Melody cell sorter. Tetramer-positive T-cells from each sample were sorted into a common tube and centrifuged on 390 g for 10 min at 4°C. The supernatant was removed, and cells were loaded into a Chromium Controller and prepared for sequencing as described in the Chromium Next GEM Single Cell 5' v2 protocol (CG000330). Library was constructed according to the manufacturer's instructions as described in the Chromium Next GEM Single Cell 5' v2 protocol (CG000330). The supernatant was removed, and cells were loaded onto Chromium according to the manufacturer's instructions. Downstream processing was performed according to manufacturer's instructions (Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index), with the Feature Barcode technology for Cell Surface Protein & Immune Receptor Mapping) (10x Genomics, USA). Gene expression (RNA), antibody-derived-tag (ADT), and hashtag-oligonucleotide (HTO) libraries were sequenced at Novogene using NovaSeq 150 paired-end.