Targeted enrichment of B cell Igs from Peripheral Blood Mononuclear Cells (PBMCs) pooled from BALB/c mice, purchased from CMLabs (catalog number: 12121912). Cells were ~77.5% 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 5’ gene expression (vdj_v1_mm_balbc_pbmc_5gex) and TCR (vdj_v1_mm_balbc_pbmc_t) specific libraries.
Libraries were prepared following the Single Cell V(D)J Reagent Kits User Guide (CG000086 Rev G).
- ~17,500 cells loaded
- 4,975 expected Ig-expressing cells recovered (based on Cd79a/Cd79b expressing fraction in the gene expression data)
- 4,852 cells detected with either single IgH or IgL/IgK or both
- 4,267 cells with productive V-J spanning (IgH, IgL/IgK) pair
- Sequenced on an Illumina NovaSeq 6000 with NovaSeq software v1.4 and with ~14,000 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