Mouse splenocytes from BALB/c mice were obtained by 10x Genomics from iQ Biosciences. 4 million cells were stained with a mixture of 12 TotalSeq™-B anti-mouse antibodies (CD3, CD4, CD8a, CD11b, CD14, CD19, CD25, CD44, CD62L, CD127, CD279, TIGIT) and 4 isotype controls (Rat IgG2b, κ isotype control; Mouse IgG1, κ isotype control; Rat IgG2a, κ isotype control; Rat IgG1, κ isotype control) following the demonstrated protocol Cell Surface Protein Labeling for Chromium Fixed RNA Profiling for Singleplex Samples with Feature Barcode technology (CG000529, Rev A), with the minor modification that the staining was done in a total of 200μL. 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, Rev B).
The Fixed RNA Gene Expression and Cell Surface Protein libraries were generated as described in the Chromium Fixed RNA Profiling for Singleplexed Samples with Feature Barcode technology for Cell Surface Protein User Guide (CG000477, Rev C) and sequenced on an Illumina NovaSeq6000 with approximately 67k read pairs per cell.
- 7,300 cells detected
- Paired-end, dual indexing: 28 cycles Read 1, 10 cycles i7, 10 cycles i5, 90 cycles Read 2