Human peripheral blood mononuclear cells (PBMCs) of a healthy male donor aged 27 were obtained by 10x Genomics from AllCells.
Gene Expression, Feature Barcode, BCR Enriched, and TCR Enriched libraries were generated from 1,690 cells (91% viable, expected cell recovery 922) as described in the Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) User Guide with Feature Barcode technology for Cell Surface Protein and Immune Receptor Mapping (CG000330 Rev A) and sequenced on an Illumina NovaSeq 6000.
Sequencing Configuration:
- Gene Expression: Paired-end (26X90), dual-indexed sequencing. Read 1: 26 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 90 cycles
- BCR Enriched: Paired-end (26X90), dual-indexed sequencing. Read 1: 26 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 90 cycles
- TCR Enriched: Paired-end (26X90), dual-indexed sequencing. Read 1: 26 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 90 cycles
- Feature Barcoding: Paired-end (26X90), dual-indexed sequencing. Read 1: 26 cycles, i7 index: 10 cycles, i5 index: 10 cycles, Read 2: 90 cycles
Analysis Settings:
- Gene Expression: --expect-cells=1000