Gene expression in the pancreas of 12 week old Non-Obese Diabetic (NOD) mice vs. healthy age-matched NOD.B10 control mice

Gene expression in the pancreas of NOD vs. NOD.B10 mice at 12 weeks of age The pancreas of 12 week old NOD and NOD.B10 mice were isolated and homogenized in Trizol reagent (Invitrogen; 1 ml/100 mg tissue). 0.2 ml of chloroform was added per ml Trizol. Samples were shaken vigorously and centrifuged at 12,000xg at 4°C. The clear aqueous phase was combined with an equal volume of 70% ethanol, and bound to a RNeasy mini column (Qiagen). RNA was extracted according to manufacturer’s instructions and RNA quality was assessed using the Agilent 2100 Bioanalyzer and the RNA 6000 Nano Reagent Kit (Agilent). Total RNA (1ug) extracted from the pancreas of NOD mice were labelled with Cy5 and run individually against a pool of Cy3-labeled total RNA (1 ug) isolated from the pancreas of age-matched NOD.B10 mice (pool of 8 animals). Microarrays were performed using the Whole Mouse Genome Microarray Kit, 4×44K 2-color arrays (Agilent Technologies).