title: GSM8017509: Islets, WT mice, scRNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: The pancreatic islets were isolated from 8-12-week-old male C57BL/6J mice. After euthanasia, the pancreas was perfused by injection of 2-4 mL digestion buffer (Hanks' Balanced Salt Solution containing type IV collagenase into the common bile duct. The pancreas was then dissected and then incubated at 37°C for 10 minutes followed by gently shaking for 3-6 minutes. Density gradient centrifugation was performed, and individual islets with high purity were hand-picked under a dissecting microscope. To prepare the single-cell suspension, the purified islets were digested with 0.25% trypsin-EDTA and filtered through a 40 μm cell strainer. Cell viability was evaluated by trypan blue staining, followed by microscopy to ensure that cell viability was greater than 90%, and over 90% of cells were singlets. Following preparation of islet single-cell suspension, the single cells were loaded onto 3' library chips for the Chromium Single Cell 3' Library (v3) according to the manufacturer's recommendations (10X Genomics)