Huang Yue-qin, Xu You-ming, Xue Jian-ping
Department of Biology, Huaibei Coal Industry Teachers' College, Huaibei 235000, China.
Zhongguo Zhong Yao Za Zhi. 2008 Aug;33(15):1810-3.
To extract RNA from Pinellia ternata and lay a foundation for studying the formation mechanism of P. ternata.
By modifying the method recommended by Guanidinium for extracting total RNA from plant tissues rich in phenolic and polysaccharidic compounds, a simple and convenient method for extraction of total RNA from the tubers, stems and leaves of P. ternate containing abundant polyphenols and polysaccharides was established. High concentrated p-mercaptoethanol was added in the RNA extracted buffer to remove polyphenols, phenol and chloroform were used to eliminate proteins, and isopropanol and sodium acetate were used to precipitate polysaccharides.
The A260/A230 value of RNA extracted with improved method were all over 2.0 and the values of A260/A280 were between 1.7 and 2.0. The electrophoresis bands were cleared on agarosegel and integrity of RNA was good.
The results showed that RNA obtained from the tubers, stems and leaves of P. ternate with this method had high purity and quality and could be used in molecular biological research, as DDRT-PCR and reverse Northern blotting analysis directly. This method is simple, economic, stable performance, and has a good repeatability as well as is suitable for extracting total RNA of medicinal plants with high concentrations of phenolics and polysaccharides.
从半夏中提取RNA，为研究半夏的形成机制奠定基础。
通过改进胍盐法推荐的从富含酚类和多糖类化合物的植物组织中提取总RNA的方法，建立了一种简单便捷的从富含大量多酚和多糖的半夏块茎、茎和叶中提取总RNA的方法。在RNA提取缓冲液中加入高浓度的β-巯基乙醇以去除多酚，用苯酚和氯仿去除蛋白质，并用异丙醇和醋酸钠沉淀多糖。
用改进方法提取的RNA的A260/A230值均大于2.0，A260/A280值在1.7至2.0之间。琼脂糖凝胶电泳条带清晰，RNA完整性良好。
结果表明，用该方法从半夏块茎、茎和叶中获得的RNA具有高纯度和质量，可直接用于分子生物学研究，如DDRT-PCR和反向Northern印迹分析。该方法简单、经济、性能稳定、重复性好，适用于提取富含高浓度酚类和多糖的药用植物的总RNA。
