Gene expression in the islets of 12 wk old Non-Obese Diabetic (NOD) mice and healthy control NOD.B10 mice

Gene expression in the islets of NOD and NOD.B10 mice at 12 weeks of age, prior to the onset of destructive insulitis. The pancreata of NOD and NOD.B10 mice (n=6) were frozen and islets were isolated by laser capture microdissection. Cryosections (8 um) were cut and stained using the Arcturus HistoGene Frozen Section Staining kit (Applied Biosystems) and laser dissection was performed using the Leica AS LMD and the Leica IM 1000 Image Manager Basic software. Sections from at least 40 individual islets were collected and RNA was extracted using the RNeasy micro kit (Qiagen). RNA quality was assessed using the Agilent 2100 Bioanalyzer and the RNA 6000 Pico Reagent Kit (Agilent). Samples were preamplified using the TrueLabeling-PicoAMP kit (SA Biosciences), post-labeled with Cy5 and run against a Cy3-labeled mouse Universal RNA control (SA Biosciences). Microarrays were performed using the Whole Mouse Genome Microarray Kit, 4x44K 2-color arrays (Agilent Technologies).本数据集包含12周龄、尚未发生破坏性胰岛炎的非肥胖糖尿病（NOD）小鼠及NOD.B10小鼠的胰岛基因表达数据。实验中取各6只NOD及NOD.B10小鼠的胰腺进行冷冻保存，随后通过激光捕获显微切割技术分离胰岛。将冰冻组织切成厚度为8微米的切片，使用Arcturus HistoGene冰冻切片染色试剂盒（Applied Biosystems）进行染色，并借助徕卡AS LMD激光显微切割系统与徕卡IM 1000图像管理基础软件完成激光切割操作。收集至少40个独立胰岛的切片样本，使用RNeasy微量试剂盒（Qiagen）提取RNA。采用安捷伦2100生物分析仪及RNA 6000 Pico试剂试剂盒（Agilent）对RNA质量进行评估。利用SA Biosciences的TrueLabeling-PicoAMP试剂盒完成样本预扩增，以Cy5对样本进行标记后，与经Cy3标记的小鼠通用RNA对照样本（SA Biosciences）进行杂交分析。最终采用安捷伦科技（Agilent Technologies）的全小鼠基因组微阵列试剂盒（4×44K双色微阵列）完成微阵列检测。