Human bone marrow aspirates from a healthy male aged 60-65 were obtained by 10x Genomics from StemExpress. Mononuclear cells were isolated, aand cells were labeled with TotalSeq™-C Human Universal Cocktail, v1.0 (BioLegend) and viable cells were identified by 7AAD staining and sorted via FACS.
Gene Expression, Cell Surface Protein/Immune Receptor Mapping, BCR Amploified, and TCR Amplifield libraries were generated from ~33,000 cells (17,645 cells recovered) as described in the Chromium Next GEM Single Cell 5' HT Reagent Kits v2 User Guide with Feature Barcode technology for Cell Surface Protein and Immune Receptor Mapping (CG000424 Rev A) using the Chromium X. Libraries were sequenced on an Illumina NovaSeq 6000 to a read depth of approximately 38,000 mean reads per cell for Gene expression, 12,000 mean reads per cell for TCR Amplified, 70,000 mean reads per cell for BCR Amplified, and 5,000 mean reads per cell for Cell Surface Protein/Immune Receptor Mapping 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.