Transcription profiling of aortic samples from newborn and 6 week old C57/black6 wild type mice

Aortic samples ranging from the aortic valve to the diaphragm were harvested from 15 newborn and 15 six-week old C57/black6 wildtype mice. Frozen samples of 5 animals each were pulverized and directly dissolved in RNA pure (PeqLab) to get three samples for each group. Total RNA was isolated using phenol/chloroform extraction. For gene-expression analysis, 500 ng total RNA of each RNA sample was labeled using the Agilent single-color Quick-Amp Labeling Kit and hybridized on Agilent Whole Mouse Genome Microarrays (4x44K). For analysis of microRNA, 200 ng total RNA of each sample was labeled using the Agilent miRNA Labeling Reagent and Hyb Kit, and hybridized on Agilent Mouse miRNA Microarrays (8x15K) according to the manufacturer's instructions.本研究从15只新生及15只6周龄的C57/BL6野生型小鼠体内采集了覆盖主动脉瓣至膈肌区域的主动脉样本。新生组与6周龄组每组各取5只动物的冷冻样本进行匀浆破碎，随后直接溶解于RNApure（PeqLab）试剂中，最终每组获得3份样本。采用酚-氯仿抽提法提取总RNA。针对基因表达分析，取每份RNA样本的500 ng总RNA，使用安捷伦单色Quick-Amp标记试剂盒进行标记，随后在安捷伦全小鼠基因组微阵列（4x44K）上完成杂交。针对微小RNA（microRNA）分析，取每份样本的200 ng总RNA，使用安捷伦微小RNA标记试剂与杂交试剂盒进行标记，并按照制造商说明书在安捷伦小鼠微小RNA微阵列（8x15K）上完成杂交。