BALB/c mouse PBMCs cells were obtained by 10x Genomics from C&M Lab Pro and CD19+ B cells were isolated by immunomagnetic selection.
Gene Expression and TCR Enriched libraries were generated from 1,670 cells (61% viable, expected cell recovery 610) as described in the Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) User Guide (CG000331 Rev A) and sequenced on an Illumina NovaSeq 6000.
These libraries were analyzed using the new Cell Ranger multi pipeline, which combines Gene Expression, Feature Barcode, TCR enriched, and BCR enriched libraries from the same sample. Cell Ranger multi enables consistent cell calling between gene expression and VDJ libraries.
Sequencing Configuration
- Paired-end (26X90), dual-indexed sequencing. Read 1: 26 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 90 cycles