杜鹃花叶片总RNA的改良CTAB法提取

Extraction of Total RNA in Rhododendron Leaf with Improved CTAB Method

目的研究杜鹃花叶片总RNA的提取方法。方法一种适合富含次生代谢产物的杜鹃花叶片总RNA的提取方法——改良CTAB法。采用CTAB作为去污剂,分别用氯仿和水饱和酚反复抽提以及高浓度NaCl沉淀,以去除蛋白质、多糖和次生代谢物等杂质,最后用无水乙醇沉淀获得总RNA。结果该方法不仅使所获得的总RNA完整性好,纯度高,而且操作简单、快速、成本低廉。结论该方法对杜鹃花等富含次生代谢产物的植物特别是富含多酚、多糖的材料的总RNA提取具有借鉴意义。

Objective To study the method for isolation of total RNA from Rhododendron leaf. Methods An improved CTAB method was presented here to be suitable for isolation of total RNA from Rhododendron leaf , a plant which contained abundant secondary metabolites. Using chemical CTAB as detergent in the extraction buffer, the absolute ethanol precipitation of RNA was performed finally after removing of proteins, polysaccharides, and secondary metabolites by subsequent chloroform, H2O - saturated phenol extraction and high concentration NaCl precipitation, respectively. Results The yielded total RNA had integrality and high purity but. Conclusion The method may be applied to some difficult RNA extraction plants full of polyphenol and polysaccharides, particularly Rhododendron.