Sorted PBMCs from a CMV Seronegative Donor - 5' Gene Expression with a Panel of dCODE™ Dextramer® Reagents and TotalSeq™-C Antibodies

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

This dataset is licensed under the Creative Commons Attribution license.