Human B cells isolated from a healthy female donor 14 days post-influenza vaccination were obtained by 10x Genomics from Discovery Life Sciences.
Gene expression and BCR Amplified libraries were generated from 16,400 cells (85% viable, expected recovery 8,500 cells) 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 Cell Ranger multi pipeline, which combines Gene Expression, Cell Surface Protein/Immune Receptor Mapping, and V(D)J Amplified libraries from the same sample. Cell Ranger multi enables consistent cell calling between gene expression and V(D)J libraries.
Sequencing Configuration
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Gene Expression: Paired-end (28X91), single-indexed sequencing. Read 1: 28 cycles, i7 index: 8 cycles, i5 index: 0 cycles, Read 2: 91 cycles. Note: Read 1 requirement is 26 bp; 28 bp Read 1 is trimmed by Cell Ranger pipeline.
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BCR Amplified: Paired-end (26x150), dual-indexed sequencing. Read 1: 26 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 150 cycles.