几种提取植物DNA方法的比较.docVIP

几种提取植物DNA方法的比较 韩玉杰1,贾炜珑2,王自霞1,周小梅1 (1.山西大学生命科学与技术学院,山西太原030006;2.天津大学化学系,天津300072 摘　要:以玉米和草的叶片为材料,分别用CT AB,S DS和尿素法提取基因组DNA,通过紫外分光光度法、琼 脂糖凝胶电泳、蒽酮比色法对所提DNA的产量、质量和糖含量进行了检测比较,并对鲜样采取了不同的干 燥处理。试验结果表明,鲜样和硅胶干燥的叶片提取的DNA无降解或降解很少,45℃烘干样也能提取到 大量的高分子DNA,自然干燥样的提取效果因植物材料而异。 关键词:植物;DNA;提取;干燥 中图分类号:Q781　　　文献标识码:A　　　文章编号:1002-2481(200807-0017-03 Com par ison of M ethods i n D NA Extracti on from Pl an ts HAN Yu2jie1,J I A W ei2l ong2,W ang Zi2xia1,ZHOU Xiao2mei1 (1.School of L ife Science and Technology,Shanxi U niversity,Taiyuan030006,China; 2.D epart m ent of Che m istry,T ianjin U niversity,T ianjin300072,China Abstract:CT AB-1method,CT AB-2method,S DS method and urea method were used t o extract the genome DNA fr om corn and weed.The resulted DNA samp les of the methods were tested using UV abs or p ti on value,agar ose gel electr opho2 resis,and anthraquinone p ig ment t o compare sugar content,yield and quality of obtained DNA.CT AB was decided as the best method f or the test.Several desiccati on methods of fresh leaves were als o investigated.The results show that quality of DNA fr om silica-gel-dried leaves is al m ost the sa me as of the fresh leaves.Leaves dried at42℃was als o yielded high molecular weight DNA with less degradati on.The DNA fr om dried leaves at r oom te mperature are different fr om p lant materials.The re2 sults reveal that DNA fr om fresh and silica Gel dried leaves materials are degrading less and qualified.DNA obtained fr om CT AB-1are well-qualified,high-purity,l ow-sugar and with high yields. Key words:Plant;DNA;Extracti on;Dehydrated desiccati on 　　植物DNA的提取和纯化是植物基因工程研究的基础操作。实施分子标记、基因图谱、基因指纹图谱、基因文库构建、遗传多态性分析、DNA 重组、RAP D(随机引物多态性DNA、RF LP(限制性酶多态性、基因分离及遗传转化和鉴定等都以提取DNA为前提。 目前,对从植物组织中提取DNA方法的要求是:简便、有效、快捷以及经济。与动物组织相比,植物组织含有较多的多糖和其他次生代谢产物(酚、酯、萜、色素等,这给高质量的DNA提取带来较多的困难[1]。尤其是多糖成分是影响DNA纯度最常见的问题。如何做到既去除这些污染物又能相对保持高的产量,并且能简捷有效地提纯,一直是植物生物技术研究者探索的问题。 另外,对于大量样品的分析和需要到资源所在地收集材料,无法对新鲜样材即刻提取的,则采取适当干燥处理后,再进行干样的DNA提取,是一种简便、经济、实用的方法。 本研究以玉米和草为材料,对4种DNA的 ? 7 1 ? 山西农业科学