Mouse splenocytes from BALB/c mice were obtained by 10x Genomics from iQ Biosciences. Cells were fixed for 1 hour at room temperature following the demonstrated protocol Fixation of Cells & Nuclei for Chromium Fixed RNA Profiling (CG000478, Rev B).
The Fixed RNA Gene Expression library was generated as described in the Chromium Fixed RNA Profiling for Multiplexed Samples User Guide (CG000527, Rev C). The fixed splenocytes were run as a 16-sample multiplexed experiment, where the sample was evenly split into 16 separate hybridization reactions, each using a unique Probe Barcode (sample sub-pooling). After hybridization, cells from each hybridization were pooled in equal proportions, washed, and then run in a single GEM lane. The resulting library was sequenced on an Illumina NovaSeq6000 with approximately 29k read pairs per cell.
- Total 113,001 cells detected
- Paired-end, dual indexing: 28 cycles Read 1, 10 cycles i7, 10 cycles i5, 90 cycles Read 2