Mouse splenocytes labeled with cell surface antibodies using TotalSeq™-C Mouse Universal Cocktail v1.0

BALB/c mouse splenocytes were obtained by 10x Genomics from IQ Biosciences. Cells were stained with TotalSeq™-C Mouse Universal Cocktail, V1.0 (BioLegend, Cat# 199903) following the demonstrated protocol Cell Surface & Intracellular Protein Labeling for GEM-X Flex Gene Expression (CG000781, Rev C). After staining, cells were washed using the 2-Wash option and then fixed overnight at 4°C following the demonstrated protocol Fixation of Cells & Nuclei for GEM-X Flex Gene Expression (CG000782, Rev D).

The Gene Expression library and Protein Expression library were generated as described in the GEM-X Flex v2 with Feature Barcode technology for Protein User Guide (CG000835, Rev B). The fixed cells were processed in 4 hybridization reactions. After Probe Hybridization, Barcode Oligo Hybridization was performed using 4 unique Barcodes from Barcode Oligo Plate Set C. After hybridization, cells were washed, and then run in a single GEM lane. The resulting libraries were sequenced on an Illumina NovaSeq6000 with approximately 72k read pairs per cell for Gene Expression library and 21k read pairs per cell for Protein Expression library.

• Approximately 16,000 total cells detected, with about 4,000 cells detected per sample
• Paired-end, dual indexing: 54 cycles Read 1, 10 cycles i7, 10 cycles i5, 60 cycles Read 2

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