Targeted enrichment of TCRs from Peripheral Blood Mononuclear Cells (PBMCs) pooled from C57BL/6 mice, purchased from CMLabs (catalog number: 11121911). Cells were ~72% 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 (vdj_v1_mm_c57bl6_pbmc_5gex) and B cell Ig (vdj_v1_mm_c57bl6_pbmc_b) specific libraries.
Libraries were prepared following the Single Cell V(D)J Reagent Kits User Guide (CG000086 Rev G).
- ~17,500 cells loaded
- 1,492 expected TCR-expressing cells recovered (based on Cd3d/Cd3e expressing fraction in the gene expression data)
- 1,396 cells detected with either single TRA or TRB or both
- 916 cells with productive V-J spanning (TRA, TRB) pair
- Sequenced on an Illumina NovaSeq 6000 with NovaSeq software v1.4 and with ~49,000 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