GSM9410200: K-AP 14 days, mKate2+ epithelial cells, replicate 2; Mus musculus; RNA-Seq
The FixNCut method was used to preserve pancreatic cells from harvested tissue. Fresh mouse pancreas tissue (~3×3 mm) was fixed in 1 mg/mL DSP (dithiobis(succinimidyl propionate)) at room temperature for 30 min and quenched with Tris-HCl. Tissue was dissociated with Liberase TM (200 µg/mL, 37 °C, 30 min), filtered (70 µm), washed in PBS + 1% BSA, and resuspended with RNase inhibitor. Single-cell suspensions were filtered (40 µm), counted, and loaded onto the 10x Chromium controller targeting ~10,000 cells. Single-cell 3′ gene expression libraries were generated using the 10x Genomics Chromium Single Cell 3′ Reagent Kit v3.1 according to the manufacturer's instructions. Libraries were sequenced on Illumina platforms using paired-end 100-cycle chemistry.