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 TCR enriched and Ig enriched libraries, following the Chromium Single Cell V(D)J Reagent Kits User Guide with Feature Barcoding technology for Cell Surface Protein (CG000186).
The libraries were sequenced on an Illumina NovaSeq 6000 (NovaSeq software v1.6).
V(D)J Enriched Libraries
TCR Libraries
- Sequencing Depth:
TCR Library - 20,183 read pairs per cell
- Sequencing Coverage:
Read 1 - 150 bp i7 Sample Index - 8 bp i5 Sample Index - 0 Read 2 - 150 bp
The key cell metrics were:
- Cells detected - 3,928
- Reads mapped to any V(D)J gene - 80.7%
- Cells with productive V-J spanning pair - 85.6%
Ig Libraries
- Sequencing Depth: Ig Library - 17,455 read pairs per cell
- Sequencing Coverage: Read 1 - 150 bp i7 Sample Index - 8 bp i5 Sample Index - 0 Read 2 - 150 bp
The key cell metrics were:
- Cells detected - 775
- Reads mapped to any V(D)J gene - 82.9%
- Cells with productive V-J spanning pair - 94.2%