番茄果实总RNA提取方法的优化

An Improved Method for Extracting RNA from Tomato Fruit

针对番茄等果实的特殊性,对商品化Trizol试剂盒提取总RNA的方法进行适当的改良:液氮研磨后加入预处理液除去果实中大部分糖类等次级代谢物质,裂解时加入β-巯基乙醇抑制酚类等物质的氧化,从富含多糖、多酚的番茄、枣果肉中成功提取总RNA。与其它方法相比,该改良方法具有操作步骤简单、快速等优点。利用RT-PCR技术,从所提取RNA中成功克隆出ζ-胡萝卜素脱氢酶基因片段,进一步表明其质量可以完全满足分子生物学研究的要求。

Extraction of high quality RNA from fruit flesh is particularly difficult due to large amount of polysaccharides,polyphenol and other secondary metabolites.A simple procedure for RNA isolation from tomato and Chinese date fruits was designed.The commercial Trizol kit protoco was improved by using pretreatment solution after grinding in liquid N2 to remove most of secondary metabolites,and adding β-mercaptoethanol into the extraction solution to prevent polyphenolic compounds from oxidizing.Compared with other methods,the procedure was simple and rapid.RT-PCR analysis of zeta-carotene desaturase gene also demonstrated that the isolated RNA samples were in high quality.