杜仲树皮总RNA提取方法的比较  

Comparison of Extraction Methods of Total RNA from Eucommia Ulmoides Oliv

作　　者：丁勇[1] 黎燕红[1] 王汉斌[1] 龚秀会[1] 刘小烛[1] 

机构地区：[1]西南林业大学西南山地森林资源保育与利用教育部重点实验室,云南昆明650224

出　　处：《安徽农业科学》2011年第6期3255-3257,共3页Journal of Anhui Agricultural Sciences

基　　金：西南地区生物多样性保育国家林业局重点实验室开放基金(KL200704);云南省教育厅科研基金(2010Y295)

摘　　要：[目的]从杜仲树皮中获得高质量的总RNA。为开展杜仲后续mRNA分离、杜仲抗真菌蛋白基因克隆、及杜仲树皮cDNA文库构建等研究奠定基础。[方法]以杜仲树皮为材料,分别采用改良CTAB-LiCl法、RNApure Plant Kit法和RNAiso Plus法进行总RNA提取,用琼脂糖甲醛变性凝胶电泳检测总RNA的完整性,用紫外分光光度计检测总RNA的纯度和得率。[结果]改良CTAB-LiCl法和RNA-pure PlantKit法提取的杜仲树皮总RNA纯度和完整性较高,A260/A280值均在1.800~2.000,28和18 SrRNA条带清晰,RNA得率较高;而RNAiso Plus法提取的杜仲树皮总RNA的纯度和完整性较低,A260/A280值为1.652,28S和18S rRNA条带存在降解和弥散现象,RNA得率低。[结论]改良CTAB-LiCl法和RNApure Plant Kit法抽提获得的杜仲树皮总RNA质量较高,能满足后续RT-PCR和RACE等试验的要求,这为成功克隆杜仲相关基因奠定了基础。关键词杜仲树皮;RNA提取;方法;[Objective] To extract high quality total RNA from the bark of Eucommia ulmoides Oliv and lay foundation for the isolation of mRNA,cloning of anti-fungi protein gene and library construction of cDNA.[Method]Total RNA from the bark of Eucommia ulmoides Oliv was extracted by modified CTAB-LiCl method,RNApure Plant Kit method and RNAiso Plus method respectively.The quality of total RNA was examined through ultraviolet spectrophotometer and formaldehyde denaturalization agarose gel electrophoresis.[Result]The results showed that total RNA extracted by modified CTAB-LiCl method and RNApure Plant Kit method had higher purity,integrity and yield rate,the ratio of A260/ A280was in the range of 1.8～2.0,the bands of 28S and 18SrRNA were clear;while the quality of total RNA obtained by RNAiso Plus method was poor,the ratio of A260/A280was 1.652,and the band of RNA was diffusion,which suggested the total RNA was degraded during the extraction process.[Conclusion]The results showed that total RNA from the bark of Eucommia ulmoides Oliv extracted by modified CTAB-LiCl method and RNApure Plant Kit method was of high purity and quality,which could meet the requirement of RT-PCR and RACE etc.experiments as well as lay the foundation for cloning genes from Eucommia ulmoides Oliv.

关 键 词：杜仲树皮 RNA提取 方法 比较 

分 类 号：S184[农业科学—农业基础科学]