甘草RNA提取方法研究

The Methods of RNA Extraction of Radix Glycyrrhizae

目的从甘草不同组织中提取高质量的RNA,为进一步合成双链cDNA,构建甘草cDNA文库奠定基础。方法采用几种常用方法(高盐溶液法、LiCl沉淀法、CTAB法和SDS法)提取甘草不同组织的RNA,并以1%琼脂糖凝胶电泳和A260/A280,A260/A230的值作为指标。结果LiCl沉淀法对于甘草不同组织中的RNA具有更好的提取效果,条带清晰,完整性较好,且A260/A280和A260/A230值均接近于2.0,根部组织产量达0.220μg/mg,叶片组织产量达0.275μg/mg。结论采用LiCl沉淀法提取甘草不同组织中的RNA具有更好的效果,适于从富含多糖的植物组织中提取RNA。

Objective To isolate the high quality RNA from different tissues of Radix Glycyrrhizae efficiently so as to lay a foundation to synthesize the cDNA and construct the cDNA library. Methods Total RNA was isolated from different tissues of Radix Glycyrrhizae by some familiar ways, such as high - salt solution, lithium chloride sedimentation, CTAB and SDS. Use 1% agarose gel electrophoresis and UV numerical value of A260/A280 and A260/A280 as the standard. Results The method of lithium chloride sedimentation had the best effect to extract total RNA from different tissues, the bands were clear on agarose gel and the gritty of RNA was good. The ratios of total RNA A260/A280 and A260/A280 were close to 2.0, respectively. The yield of the root was 0. 220 μg/mg fresh weight, and the leaf was up to 0.275μg/mg fresh weight. Conclusion The method of lithium chloride sedimentation has the best effect to extract total RNA of different tissues, and this method is also applicable to the plants with much amylase.