紫山药组织RNA的提取方法

A New Protocol for Total RNA Isolation from Dioscorea alata Lirm.sp

针对紫山药富含次生代谢物质的特点，探索出一种可有效去除其酚类、多糖和 DNA等杂质的紫山药总RNA提取方法。在 SDS法的基础上，于粉碎材料时，用水不溶性聚乙烯吡咯烷酮（PVP）防止酚类物质氧化，同时以高盐溶液去除多糖污染。利用这一方法提取的紫山药多个组织的总 RNA，经琼脂糖电泳鉴定，可见清晰的18和28 S RNA的2条带型，表明 RNA完整，无弥散或降解；经过紫外吸收分析，A260／A380、A260／A2300分别在1．86～2．01，2．17～2．65，纯度良好，所提取的总 RNA质量可以满足进一步分子生物学研究的要求。

Based on the characteristic of Dioscorea alata Lirm.sp,mainly for its richness in polysaccharide and polyphenols,a modified protocol for RNA isolation from D.alata was developed.This new protocol could effectively remove polyphenols,polysaccharides and DNA from total RNA.In the modification meth-od based on the SDS,insoluble polyvinylpyrrolidone were used to prevent polyphenols from being oxidized in the grinding time,and polysaccharides were removed with high-salt solution.The total RNA of D.alata extracted by this method showed fine integrity and clear bands of 28 S and 18 S rRNA,indicating that the total RNA was intact.The values of A260/A280 and A260/A230 were1.86~2.01 and 2.17~2.65,respectively, which meant that the RNA could meet the demand of molecular biology experiment.