title: GSM8793169: KC2; Mus musculus; RNA-Seq
library_name: GSM8793169
library_construction_protocol: Harvested pancreata were rinsed with PBS, minced into 1 mm³ pieces on ice, and transferred to a 20 mL digestion buffer containingCollagenase P (20 mg, Roche, 11213857001), DNase I (200uL 10mg/mL, Roche 10104159001), Fetal Bovine Serum (400 µL), and Rho Kinase inhibitor (Y-27632) (20 µL, Cayman Chemicals,  10005583). The mixture was incubated for 75 minutes at 37°C with continuous rotation. Samples were then passed through a 70 μm cell strainer (Cell Treat,  229483), and the tube was rinsed with 20 mL of ice-cold PBS. Following centrifugation at 700 rpm for 5 minutes at 4°C, the pellet was resuspended in 1 mL of PBS containing 20 mM HEPES, 2 mM EDTA, 5mM MgCl2, 1% FBS, 50ug/mL DNase I, and Rho Kinase inhibitor (Y-27632). Cells were stained with DAPI, and viable cells were sorted using the BC Influx cell sorter. The viability of the cell samples was confirmed to be greater than 80%, and the cells were resuspended in PBS containing 10% FBS at a concentration of 1,000 cells/µL. The resulting single-cell suspension was used for scRNA-seq. We aimed to profile 10,000 cells per library. All libraries were sequenced on the NovaSeq6000 (Illumina, San Diego, California, USA) or DNBSEQ-G400 (MGI Tech, Shenzhen, China)