Peripheral blood mononuclear cells (PBMCs) from a CMV seronegative donor, were obtained from Astarte (donor 396, catalog #: 1001-3894AP18). 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”.
The PBMCs were sorted for CD4-/CD8+/dextramer+ T cells.
Cells were 87% viable by Trypan blue stain.
The same cDNA was used to generate TCR (vdj_v1_hs_sorted_cmvneg_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
- 1,547 cells detected
- 1,502 median genes per cell and 4,787 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 ~51,000 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