Human peripheral blood mononuclear cells (PBMCs) of a healthy donor were obtained from AllCells (part number: PB001). The cells were 87% viable (trypan blue stain). The cells were labeled with a panel of 14 TotalSeq™-C antibodies and 3 isotype controls (BioLegend).
Amplified cDNA from the labeled cells were used to generate 5’ gene expression, cell surface protein, TCR enriched, and Ig enriched libraries, following the Chromium Single Cell V(D)J Reagent Kits User Guide with Feature Barcode technology for Cell Surface Protein (CG000186).
The libraries were sequenced on an Illumina NovaSeq 6000 (NovaSeq software v1.6).
5’ Gene Expression + Cell Surface Protein Libraries
- Sequencing Depth: 5’ Gene Expression Library - 89,433 read pairs per cell Cell Surface Protein Library - 20,959 read pairs per cell
- Sequencing Coverage: Read 1 - 28 bp i7 Sample Index - 8 bp i5 Sample Index - 0 Read 2 - 91 bp
The key cell metrics were:
- Cells detected - 7,231
- UMI counts - 4,279
- Median genes per cell - 1,384
V(D)J Enriched Libraries
TCR Libraries
- Sequencing Depth: TCR Library - 21,233 read pairs per cell
- Sequencing Coverage: Read 1 - 26 bp i7 Sample Index - 8 bp i5 Sample Index - 0 Read 2 - 91 bp
The key cell metrics were:
- Cells detected - 3,925
- Reads mapped to any V(D)J gene - 84.3%
- Cells with productive V-J spanning pair - 85.5%
Ig Libraries
- Sequencing Depth: Ig Library - 18,143 read pairs per cell
- Sequencing Coverage: Read 1 - 26 bp i7 Sample Index - 8 bp i5 Sample Index - 0 Read 2 - 91 bp
The key cell metrics were:
- Cells detected - 772
- Reads mapped to any V(D)J gene - 83.1%
- Cells with productive V-J spanning pair - 93.4%