Peripheral blood mononuclear cells (PBMCs) from a CMV seropositive donor were obtained through a collaboration. Cells were stained with a panel of 12 dCODE™ Dextramer® reagents (Immudex) and 17 TotalSeq™-C antibodies (BioLegend). See details in the Application Note “Redefining Cellular Phenotyping: Comprehensive Characterization and Resolution of the Antigen-Specific T Cell Response”.
Cells were 81% viable by Trypan blue stain.
The PBMCs were sorted for CD4-/CD8+/dextramer+ T cells.
The same cDNA was used to generate TCR (vdj_v1_hs_sorted_cmvpos_t) specific libraries.
Libraries were prepared following the Single Cell V(D)J Reagent Kits User Guide (CG000186 RevA).
- All cells collected from the sort were loaded, due to the expected low cell number an accurate count of loaded cells could not be performed
- 2,043 cells detected
- 1,308 median genes per cell and 4,580 median UMI counts per cell detected
- Gene expression data was sequenced on an Illumina NovaSeq (NovaSeq Software v1.6) with ~55,000 read pairs per cell; cell surface protein data was sequenced on a HiSeq4000 with ~21,000 read pairs per cell
- 28bp read1 (including 16bp Chromium barcode and 10bp UMI), 91bp read2 (transcript), and 8bp I7 sample barcode read configuration for gene expression and cell surface protein data