Flash-frozen tissues from C57BL/6 mice were obtained by 10x Genomics from BioIVT:
- Eye (pair, 40 mg)
- Ovary (pair, 28 mg)
- Stomach (45 mg)
- Small Intestine (69 mg)
Frozen tissues were processed following the demonstrated protocol Tissue Fixation & Dissociation for Chromium Fixed RNA Profiling (CG000553) and fixed for 24h at 4°C. After fixation, the fixed tissue pieces were roughly divided into two aliquots, one of which was dissociated using the gentleMACS Octo Dissociator.
The Fixed RNA Gene Expression library was generated as described in the Chromium Fixed RNA Profiling for Multiplexed Samples User Guide (CG000527, Rev C). Samples were run as a 4-sample multiplexed experiment, in which each sample was hybridized with a unique Probe Barcode in a separate hybridization reaction. After hybridization, samples 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 30k read pairs per cell.
- Total 42,704 cells detected (Eye: 13,205; Ovary: 10,545; Stomach: 10,038; Small Intestine: 8,916)
- Paired-end, dual indexing: 28 cycles Read 1, 10 cycles i7, 10 cycles i5, 90 cycles Read 2
Brightfield and propidium iodide fluorescence images of the resulting cell suspensions after dissociation are provided, as well as images of the final pooled sample (also shown below).
| Brightfield | Fluorescence |
|---|---|
 |  |