Non-small cell lung cancer (NSCLC) dissociated tumor cells from 7 donors were obtained from Discovery Life Sciences. Cells were labeled with TotalSeq™-B Human TBNK Cocktail (BioLegend). Each donor was CellPlexed and pooled at equal proportions. Viable cells in the pool were identified by 7AAD staining and sorted via FACS.
Gene expression and CellPlex libraries were generated as described in the Chromium Next GEM Single Cell 3' HT Reagent Kits v3.1 User Guide with Feature Barcode technology for Cell Surface Protein and Cell Multiplexing (CG000420 Rev A) using the Chromium X. Libraries were sequenced on an Illumina NovaSeq 6000 to a read depth of approximately 50,000 mean reads per cell for gene expression and 35,000 mean reads per cell for CellPlex.
Paired-end, dual indexing Read 1: 28 cycles (16 bp barcode, 12 bp UMI) i5 index: 10 cycles (sample index) i7 index: 10 cycles (sample index) Read 2: 90 cycles (transcript).
Run with --expect-cells= 40000.