40k Mixture of Nuclei Isolated from 4 Mouse Tissues, Multiplexed Samples, 4 Probe Barcodes

Flash-frozen tissues from C57BL/6 mice were obtained by 10x Genomics from BioIVT:

• Eye (pair, 47 mg)
• Ovary (pair, 23 mg)
• Stomach (77 mg)
• Small Intestine (52 mg)

Nuclei were isolated from following the Chromium Nuclei Isolation Reagent Kits Sample Prep User Guide (CG000505) and then 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). 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 41k read pairs per cell.

• Total 35,661 cells detected (Eye: 8,568; Ovary: 9,862; Stomach: 9,996; Small Intestine: 7,235)
• 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 nuclei suspensions after fixation are provided, as well as images of the final pooled sample (also shown below).

Brightfield	Fluorescence

This dataset is licensed under the Creative Commons Attribution license.