光皮桦基因组DNA提取方法比较  

Comparison of methods of extracting genomic DNA from Betula luminifera

作　　者：谢一青[1] 李志真[1] 黄儒珠[2] 肖祥希[1] 王志洁[1] 

机构地区：[1]福建省林业科学研究院,福建福州350012 [2]福建师范大学生命科学学院,福建福州350007

出　　处：《浙江林学院学报》2006年第6期664-668,共5页Journal of Zhejiang Forestry College

基　　金：福建省林业厅重大科学技术攻关项目(200306)

摘　　要：为从富含次生代谢物的光皮桦Betula luminifera嫩叶中获得高质量DNA,设计了5种先提核再提DNA的CTAB法Ⅰ,CTAB法Ⅱ,CTAB法Ⅲ,CTAB法Ⅳ及改进的SDS法,并以常规CTAB法为对照。用不同的方法就光皮桦基因组DNA进行了提取,采用琼脂糖凝胶电泳检测、A260/A280值测定、限制性内切酶处理和PCR扩增等方法对所提的DNA进行了比较分析。结果表明:实验设计的5种改进方法提取的DNA质量要比常规法的好,但提取的效果差异较大。其中改进的CTAB法Ⅰ是提取光皮桦基因组DNA的最佳方法;PCR扩增结果表明,不同的DNA提取方法会影响PCR带型的变化。Six methods including basic CTAB method, modified CTAB methods as CTABⅠ, CTAB Ⅱ , CTAB Ⅲ, CTAB Ⅳ, and improved SDS methods were used to extract high-quality DNA from the young leaves of Betula luminifera. The DNA samples obtained by the above methods were tested by agarose-gel electrophoresis, restricted enzyme digests and PCR. Results showed that the modified methods were better than the basic CTAB method in terms of the quality of total DNA, but the effects of DNA extraction differed greatly. Among them, the modified CTAB Ⅰ method was the best for extracting B. luminifera DNA from leaves. The comparisons of PCR patterns of genomic DNA extracted with different methods showed that different extracting methods would affect the changes of PCR patterns. [Ch, 3 fig. 1 tab. 13 ref. ]

关 键 词：植物学 光皮桦 DNA提取 CTAB法 SDS法 顽拗类植物 

分 类 号：S718.43[农业科学—林学]