改进TRIpure法提取小鼠肝脏组织总RNA.pdfVIP

实用预防医学 2008年2月 第 15卷 第 1期 Practical Preventive Medicine，Feb．2008，Vol 15，No．1 19 文章编号：1006—3110(2008)叭一0叭9—03 【论 著 】 改进TRIpure法提取小鼠肝脏组织总RNA 易尚辉 ，易银沙 ，周 鑫 ，袁炳秋 ，许 丹 ，吕 媛 摘要： 目的 改进从新鲜动物组织中提取、鉴定纯度及完整性较高总RNA的技术方法。 方法 参考1987年 Chomczynski P等人提出的RNA提取方法，对组织RNA提取的操作步骤进行调整改进以获得稳定良好的RNA，并对常规 琼脂糖凝胶电泳做适当改进用于鉴定RNA的质量。 结果 用该法提取的总RNA进行电泳后，可见28 s、18 s、5 s三 条清晰的RNA条带，其中28 s条带的亮度约为18 s条带的2倍，用紫外分光光度计检测RNA的纯度，0D260／()D280值 为1．8～2．0，表明RNA没有被蛋白质或苯酚污染。这些结果显示RNA未降解并有较好的完整性，全部操作过程可在2 h 内完成。 结论 用改进方法提取的新鲜组织RNA质量稳定，电泳鉴定快速可靠。 关键词： TRIpure；总RNA；提取；肝组织 中图分类号：R一332 文献标识码：A Isolation of Total RNA from Rat Liver Tissue by Improved Trizol Method Yl Shang—hui，Yl yin—sha，ZHOU Xin，et a1．(Medical College of Hunan Normal University，Changsha 410006，Hunan，China) Abstract： Objective To improve a technique that is employed to isolate and identify high purity and integrity of total RNA extracted frOm fresh animal tissue． Methods Referring to the RNA isolation method proposed by ChomczynskiP (1 987)，we improved the protocol of RNA isolation．Furthermore，we adjusted the conventional agarose gel eIectrophoresis to be better used to identify the RNA quality． Results When total RNA，extracted by use of the method，was used for elec— trophoresis，it clearly showed three RNA bands of 28S，18S，and 5S．The brightness of 28S RNA was twice as light as that of 1 8S RNA in the eIectrODhOresis map．Purity analysis of total RNA was conducted with UV spectrophotometer．The value of OD260／OD280 varied from 1．8 to 2．0．which indicated that the RNA was not contaminated by proteins or pheno1．The results indicated that the total RNA was complete and not degraded；and the entire procedure can be completed within 2 hrs． Con· clusions The quality of RNA isolated frOm fresh tiss