适于基因芯片实验的小鼠脑组织RNA提取方法的改进  

Optimization of RNA extraction from mouse brain for microarray analysis

作　　者：隋雷鸣[1] 赵焕英[1] 巩云霞 付文卓[1] 王俐勇[1] 

机构地区：[1]首都医科大学医学实验与测试中心,北京100069 [2]山东省莒县人民医院急诊科

出　　处：《山西医科大学学报》2014年第10期992-995,共4页Journal of Shanxi Medical University

基　　金：北京市自然科学基金青年基金资助项目(7144188);北京市教育委员会科技计划面上基金资助项目(KM201410025004);首都医科大学科研基金资助项目(2014JS06;2014JS08)

摘　　要：目的比较Trizol与两种常用提取试剂盒对小鼠脑组织RNA提取质量的差异,并进行方法改进,以期找到适合于基因芯片的高质量组织RNA提取方法。方法以小鼠脑组织为材料,Trizol试剂、Ambion和天根组织RNA提取试剂盒提取总RNA,并对试剂盒提取方法进行改良,检测改良前后RNA浓度、纯度和完整性以及反转录成cRNA的浓度。结果与改良前相比,试剂盒经方法改良后提取的RNA产量均有提高;Ambion试剂盒提取的RNA纯度优于Trizol和天根试剂盒。结论Ambion试剂盒经方法改良后RNA提取质量优于其他方法,适合于基因芯片的实验要求。Objective To explore an optimal method for the extraction of high quality RNA in microarray analysis by comparing and improving Trizol and two common RNA extraction kits methods for RNA extraction from mouse brain. Methods Total RNA of mouse brain was isolated with the Trizol, Ambion and Tiangen tissue RNA extraction kits, and RNA extraction kits methods were optimized. The concentration and purity of RNA and cRNA were determined by ultraviolet spectrophotometer, and the integrity was determined by agarose gel electrophoresis. Results The optimized procedure for RNA extraction displayed higher yield of total RNA. The purity of RNA with Ambion kit method was better than Trizol and Tiangen kit method. Conclusion The concentration and purity of RNA by optimized procedure of Ambion kit are better than those by other methods, and are suitable for microarray analysis.

关 键 词：RNA提取 基因芯片 小鼠 

分 类 号：Q78[生物学—分子生物学]