40k Mixture of Dissociated Tumor Cells from 3 Donors Stained with TotalSeq™-C Antibodies (Next GEM)

Dissociated Tumor Cells (DTCs) from three donors (Kidney Cancer, Clear Cell; Lung Cancer, (NSCLC) Pleomorphus Carcinoma; Breast Cancer, Invasive Ductal Carcinoma) were obtained by 10x Genomics from Discovery Life Sciences.

Between 470,000 to 800,000 cells per sample were stained with 32 TotalSeq™-C Antibodies following the demonstrated protocol Cell Surface Protein Labeling for Chromium Fixed RNA Profiling (CG000529, Rev B). 10% FBS in PBS was used for all labeling and wash buffers, as suggested for samples with lower viability. After staining, cells were washed using the 2-Wash option and then fixed for 1 hour at room temperature (20°C) following the demonstrated protocol Fixation of Cells & Nuclei for Chromium Fixed RNA Profiling (CG000478).

The Fixed RNA Gene Expression library was generated as described in the Chromium Fixed RNA Profiling - Multiplexed Samples with Feature Barcode technology for Protein using Barcode Oligo Capture User Guide (CG000673, Rev A). The fixed cells were run as a multiplexed experiment, where the Kidney Cancer, Breast Cancer, and Lung Cancer samples all used unique pairs of Probe Barcodes and Antibody Multiplexing Barcodes. After hybridization, cells were washed, and then run in a single GEM lane. The resulting libraries were sequenced on an Illumina NovaSeq6000 with approximately 92k read pairs per cell for Gene Expression library and 20k read pairs per cell for Protein Expression library.

• 42,120 cells detected (Kidney rep 1: 10,043; Kidney rep 2: 10,931; Lung: 10,279; Breast: 10,867)
• Paired-end, dual indexing: 28 cycles Read 1, 10 cycles i7, 10 cycles i5, 90 cycles Read 2

This dataset is licensed under the Creative Commons Attribution license.