忍冬科部分植物DNA提取方法的研究

Study on methods of total DNA extraction from some species of Caprifoliaceae

应用改进的高盐低pH法、改良高盐法、溴化十六烷基三甲基铵(CTAB)法和改良十二烷基硫酸钠(SDS)法,对七子花不同器官的总DNA进行了提取.结果显示,4种方法均有较好的提取效果,产率由大到小依次为改良SDS法、CTAB法、高盐低pH法、改良高盐法,纯度依次为改良高盐法、高盐低pH法、CTAB法、改良SDS法,其中高盐低pH法提取的综合效果最好.对七子花不同组织DNA的提取显示,产率由高到低依次是嫩叶、茎和老叶,质量以嫩叶和茎抽提的较高.以冰冻茎为材料应用筛选出的高盐低pH法对9种忍冬科植物的总DNA进行了提取,结果发现多糖、酚或胶类物质含量较高植物的DNA沉淀中有一定量的次生物质;对方法进行了少许改进,可以有效去除这些物质对DNA提取的干扰.

Four methods were used to extract genomic DNA of Heptacodiura miconioides from its dif- ferent materials. The results showed that four methods could effectively eliminate the affections of the secondary materials to genome DNA. According to the average yields of total DNA from high to low, the four methods were ranked as SDS improved method, CTAB method, high salt and low pH method and high salts improved method, and ranked as high salts improved method, high salts and low pH method, CTAB method and SDS improved method according to the total DNA purities from high to low. Compared with the four extraction methods, high salts and low pH method was the best one for total DNA extraction. The four methods were applied to extract total DNA from different organs of H. miconioldes, and the results showed that tender leaves and annual or biennial stems had a higher purity and yield. Then high salts and low pH method was applied to extract total DNA from frosted stems of nine Caprifoliaceae species. Because of the high content of polysaccharides and other secondary metabolites in some species, the method could not effectively eliminate the affections of the secondary materials in the extracted total DNA. To those questions, if the extraction method were slightly improved, it could also eliminate effectively the affections of those materials in extracted total DNA which were suitable for RAPD analysis.