GSM1580555: JY1-PA75; Mus musculus; RNA-Seq
Mice were euthanized by cervical dislocation following isoflurane anesthesia, and pancreatic islets were isolated as previously described (Ye et al., 2010b). Immediately after euthanasia, the major duodenal papilla of the mouse was blocked with a micro bulldog clamp. Ice-cold digestion solution, i.e. Hank's Balanced Salt Solution (HBSS) with 0.1 g/L Liberase TL (Roche # 05401020001, Indianapolis, IN), 0.1 g/L DNase I (Roche # 10104159001), 2.5 mM HEPES, 8 mM glucose, 0.2% BSA, pH 7.2-7.4, was inject via the common bile duct into the pancreatic duct. The inflated pancreas was transferred into a scintillation vial with digestion solution on ice. Subsequently, the vial was incubated at 37°C water bath for 25 min, and shaken vigorously to disperse the pancreas. The digested content was resuspended with ice-cold HBSS and settled on ice for 2-5 min, and supernatant was removed without disturbing the precipitate. The wash was repeated until the supernatant became clear, and the content was transferred into a 6-cm Petri dish with ice-cold HBSS plus 2.5 mM HEPES, 8 mM glucose, 0.2% BSA, pH 7.2-7.4. Under a dissection microscope, islets of Langerhans were picked with a 200-μL pipette to new dishes until free of exocrine pancreas content. The freshly isolated islets were either frozen immediately in liquid nitrogen followed by -80°C storage. Total RNA was purified from isolated pancreatic islets with RNeasy Mini Kit (Qiagen #74106) plus on-column DNase digestion. RNA libraries were prepared for sequencing using standard Illumina protocols.
GEO Accession
GSM1580555