128k Human PBMCs Stained with TotalSeq™-C Human Universal Cocktail (Next GEM)

Peripheral blood mononuclear cells (PBMCs) from a healthy donor were obtained by 10x Genomics from AllCells.

Cells were stained with TotalSeq™-C Human Universal Cocktail, V1.0 (BioLegend, Cat# 399905) following the demonstrated protocol Cell Surface Protein Labeling for Chromium Fixed RNA Profiling (CG000529, Rev B). True-Stain Monocyte Blocker (BioLegend, Cat# 426101) was added during incubation with Human TruStain FcX™ (BioLegend, Cat# 422301). 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 processed in 16 hybridization reactions using pairs of unique 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 35k read pairs per cell for Gene Expression library and 16k read pairs per cell for Protein Expression library.

• Approximately 140,000 total cells detected, with about 8,750 cells detected per sample
• 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.