“顽拗”植物春石斛基因组DNA的提取研究

Study on Extraction of Genomic DNA from Recalcitrant Plant Spring Dendrobium

[目的]提取"顽拗"植物春石斛的基因组DNA,并对其进行质量检测。[方法]采用普通CTAB法、试剂盒法和改良CTAB法提取10个春石斛品种的基因组DNA,然后分别用紫外分光光度计法、琼脂糖凝胶电泳法和荧光AFLP法对3种方法所提取的基因组DNA质量进行检验。[结果]改良CTAB法所提取春石斛基因组DNA得率较高,基本排除了春石斛体内多糖等次生代谢物质对基因组DNA质量的干扰,完全能满足AFLP等分子生物学试验对DNA质量的要求;其他2种方法所提取的基因组DNA则不能满足试验要求。[结论]改良CTAB法为较好的适合春石斛基因组DNA提取的方法。

[ Objective ] The research aimed to extract the genomic DNA from Spring Dendrobium, and examine its quality. [ Method] Three different methods, CTAB method, kit method and modified CTAB method, were chose to extract the high quality DNA from the recalcitrant spring Dendrobium. Then UV speetrophotometer method, ngarose gel electrophoresis method and fluorescent AFLP method were used to examine the quality of genomie DNA extracted by 3 extraction methods. [ Result ] The ratios of the genomic DNA extracted by the modified CTAB method were higher, and could meet the require of molecular biological experiment. But the genomic DNA extracted by the other two methods could not meet the require. [ Conclusion] The modified CTAB method is good and suitable for extracting the genomic DNA from Spring Dendrobium.