1:1 mixture of fresh frozen human HEK293T and mouse NIH3T3 cells obtained by 10x Genomics from ATCC were used in a classic experiment to demonstrate single cell behavior.
Library was generated from ~33,000 cells (18,299 cells recovered) as described in the Chromium Next GEM Single Cell 3' HT Reagent Kits v3.1 User Guide (CG000416 Rev A) using the Chromium X and sequenced on an Illumina NovaSeq 6000 to a read depth of approximately 50,000 mean reads per cell.
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=20000.