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油菜种子高质量总RNA提取的一种有效方法

An Efficient Method for Extraction of High Quality RNA from Rapeseeds

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【Author】 DING Yong ~(1))LIU Ying ~(2))YANG Yang-yang ~(1))GAN Li ~(1))(~(1))National Key Laboratory of Crop Genetic Improvement,Huazhong Agricultural University,Wuhan 430070,China;~(2))The Key Laboratory of Forest Plant Ecology Ministry of Education,Northeast Forestry University,Harbin 150040,China)

【机构】华中农业大学作物遗传改良国家重点实验室；东北林业大学森林植物生态学教育部重点实验室；华中农业大学作物遗传改良国家重点实验室武汉430070；哈尔滨150040；武汉430070；

【摘要】设计了1种从油菜种子中分离高质量总RNA的方法,有效的排除了种子中富含的多酚、多糖和蛋白质等杂质的污染。分离的总RNA经甲醛变性凝胶电泳和紫外光检测,可见28S和18S rRNA的2条清晰的带型,完整无降解,其A260/A280在1.8~2.0之间。研究结果证明,利用本方法提取的油菜种子总RNA具有很高的质量和纯度,且得率高,完全能满足后续的RT-PCR、全长基因的克隆和Northern blotting分析等分子生物学研究。更多还原

【Abstract】 A highly efficient method of extracting total RNA from rapeseed was developed.Polyphenols,polysaccharides and protein,rich in rapeseed cells,could be removed effectively.Two clear bands of 28S rRNA and 18S rRNA isolated from rapeseeds could be detected by formaldehyde denaturalization agarose gel electrophoresis.It was proved that RNA was intact without degradation.The ratio of A260/A280 is 1.8~2.0.The results showed that the RNA obtained from rapeseed with this method had high purity and quality and could be used in molecular biological research,like RT-PCR,full-length cDNA cloning and Northern blotting analysis directly.更多还原

【关键词】油菜种子； RNA提取；多酚；多糖；方法；
【Key words】 rapeseed； RNA isolation； polyphenols； polysaccharides； method；

【基金】国家自然科学基金项目(30170600);湖北省自然科学基金项目(2001ABD113)资助

【文献出处】华中农业大学学报 ,Journal of Huazhong Agricultural University(Natural Science Edition) , 编辑部邮箱 ,2006年05期

【分类号】S565.4
【被引频次】27
【下载频次】511

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