几种DNA提取方法对红树植物DNA提取效果的比较-再改稿精讲.docVIP

几种DNA提取方法对红树植物DNA提取效果的比较 丁文勇1,2陈少波1,2 **仇建标2刘伟成2(1温州医学院生命科学学院 浙江 温州 325035，2浙江省海洋水产养殖研究所浙江省近岸水域生物资源开发与保护重点实验室( 浙江 温州 325005712100 ) 摘要：红树林是发育于特殊环境的植物群落，红树植物也因其富含单宁而出名，其细胞壁含有大量的单宁和多糖等物质。单宁是一种能与蛋白质和其他化合物形成交联能力的酚化合物，核酸提取过程中，大量的单宁能与蛋白质结合，而且单宁与生物碱和多糖也可发生跟单宁与蛋白质结合相似的复合反应，而大量多糖的存在，能与DNA形成共沉淀，从而影响了核酸的纯度。以红树植物秋茄叶片为实验材料，美人蕉叶片为对照材料，采用6种不同的DNA提取方法，比较不同方法的DNA提取效果。结果表明：针对红树植物优化的CTAB法（方法6）通过PVP的除酚去多糖作用，利用CTAB提取液和氯仿-异戊醇强效除蛋白和脂类物质，结合高盐环境下异丙醇高效沉淀DNA去除多糖，最终达到彻底去除杂质，得到高纯度的基因组DNA，是6种方法中最适合用于红树植物DNA的提取方法。 关键词：红树植物；DNA提取；多糖；改进的CTAB法 Comparison of the efficiency of different DNA extraction methods on mangrove Ding Wen-yong1,2 ,Chen Shao-bo1,2, Qiu Jian-biao2, Liu Weicheng2, Xu Hai3, Wang Yiquan3 (1 School of Life Sciences, Wenzhou Medical College, Wenzhou 325015, Zhejiang, China, 2Zhejiang Mariculture Research Institute, Zhejiang Key Laboratory of Exploitation and Preservation of Coastal Bio-resource, Wenzhou 325005, Zhejiang, China, 3Colleage of Resources and Environment, Northwest A F University712100, Shanxi, China,) Abstract: Mangrove is a very special plant community in shallow tidal flat of land-sea border. It is famous for its richness in vegetable tannins and polysaccharides in its cell walls and when it is cut and oxidated it turns to be red. Tannin is a kind of phenolic compounds which can get combined with proteins and other compounds like polysaccharides by its cross-linking ability. When extracting DNA from the mangrove material, the large quantities of tannins get together with proteins and polysaccharides , and polysaccharides can also coprecipitate together with DNA, thus affecting the purity of nucleic acid. In this paper, we used 6 different methods to extract DNA from the leaves of the mangrove plant Kandelia obovata. By comparing different methods of DNA extraction, we found that: the modified CTAB method (Mothod.6 in this paper) , by using PVP and 5 mol/L NaCl to get rid of tannins and polysaccharides，together with the function of CTAB buffer, Chloroform: Isoamy