60k Human PBMCs Stained with 8 TotalSeq™-B Human Hashtags (Donor 1-4, 4 samples)

Peripheral blood mononuclear cells (PBMCs) from four diseased human donors were obtained by 10x Genomics from Sanguine Biosciences. For each donor, 250,000 cells were stained according to the manufacturer’s guidelines on antibody reconstitution and the 10x Genomics Cell Surface Protein Labeling for Single Cell RNA Sequencing Protocols with Feature Barcode technology (CG000149, Rev D). After staining, cells were washed using the Wash Protocol 2 option. Cells from the four donors were multiplexed at roughly equal proportions with two hashtags per donor, resulting in a pooled sample labeled with 8 hashtags.

Gene Expression and Cell Surface Protein libraries were generated as described in the Chromium GEM-X Single Cell 3' Reagent Kits v4 with Feature Barcode technology for Cell Surface Protein User Guide (CG000732) and sequenced on an Illumina NovaSeq 6000 with approximately 53,000 read pairs per cell for the Gene Expression library and 6,000 read pairs per cell for the Cell Surface Protein library.

Paired-end, dual indexing libraries were sequenced with the following configuration:

• 28 cycles Read 1
• 10 cycles i7
• 10 cycles i5
• 90 cycles Read 2

Libraries were analyzed using the cellranger multi pipeline.

This dataset is licensed under the Creative Commons Attribution 4.0 International (CC BY 4.0) license. 10x citation guidelines available here.