Peripheral blood mononuclear cells (PBMCs) of a healthy donor, purchased from AllCells (Catalog #: PB001), 87% viable by Trypan blue stain. PBMCs are primary cells with relatively small amounts of RNA (~1pg RNA/cell).
The same cDNA was used to generate gene expression and cell surface protein (vdj_v1_hs_pbmc2_5gex_protein) and TCR (vdj_v1_hs_pbmc2_t) specific libraries.
Libraries were prepared following the Single Cell V(D)J Reagent Kits User Guide (CG000186 RevA).
- ~17,000 total cells loaded
- 1,888 B cells expected based on CD79A/CD79B expressing fraction in 5’GEX data; 653 B cells expected based on CD19 expressing fraction in 5’GEX data
- 1,274 B cells expected based on CD19+ fraction in antibody data (CD19+ antibody cells identified by gating on UMI counts relative to IgG1 isotype control)
- 1,232 cells detected with either single IgH or IgL/IgK or both
- 91% cells with productive V-J spanning (IgH,IgL/IgK) pair
- Sequenced on an Illumina NovaSeq 6000 (NovaSeq software v1.4) with ~10,000 read pairs per cell
- 150bp read1 (including 16bp Chromium barcode, 10bp UMI, 13bp switch oligo and transcript), 150bp read2 (transcript), and 8bp I7 sample barcode read configuration