Human peripheral blood mononuclear cells (PBMCs) of a healthy male donor aged 30-35 were obtained by 10x Genomics from AllCells.
Gene expression and V(D)J libraries were generated from ~33,000 cells (18,470 cells recovered) as described in the Chromium Next GEM Single Cell 5' HT Reagent Kits v2 User Guide (CG000421) using the Chromium and sequenced on an Illumina NovaSeq 6000 to a read depth of approximately 25,000 mean reads per cell for Gene Expression, 15,000 mean reads per cell for TCR Amplified, and 25,000 mean reads per cell for BCR Amplified libraries.
Paired-end, dual indexing
- Read 1: 26 cycles (16 bp barcode, 10 bp UMI)
- i5 index: 10 cycles (sample index)
- i7 index: 10 cycles (sample index)
- Read 2: 90 cycles (transcript)
Run with --expect-cells=20000