30k Sorted DTC Multiplexed, 4 CMOs and Stained with a Panel of TotalSeq™-B Antibodies

Cells from four dissociated lung tumors (Lung Cancer (Female, Age late 60s); Squamous Cell Carcinoma (Male, Age late 80s); Adenocarcinoma (Male, Age mid 60s); NSCLC (Female, Age mid 60s)) were obtained by 10x Genomics from Discovery Life Sciences. Cells from the four dissociated lung tumors were each stained with a panel of 17 TotalSeq™-B antibodies (BioLegend) and multiplexed at equal proportions with 1 CMO per tumor, resulting in a pooled sample labeled with 4 CMOs.

Libraries were prepared following the Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 (Dual Index) with Feature Barcode technology for Cell Surface Protein and Cell Multiplexing User Guide (CG000390) and sequenced on Illumina NovaSeq 6000.

Single Cell 3’ Cell Multiplexing v3.1 Dual Index Library

• Sequencing Depth: 56,443 read pairs 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)

Pooled Multiplexed Sample - Key Metrics

• Estimated Number of Cells: 27,803
• Cells Assigned to a Sample: 22,031

Individual Multiplexed Samples - Key Metrics