五种提取马尾松基因组DNA方法的比较

Comparison of Five Methods of DNA Extraction From Pinus massoniana

对于含有大量多糖如酚、酯、萜等其它二次代谢产物的松科和杉科等针叶植物 ,要从其组织中提取高质量的基因组DNA一般都比较困难。本文以马尾松 (Pinusmassoniana)针叶为材料 ,分别采取了简易提取法、高盐沉淀法、CTAB沉淀法、Ziegenhagen法和QIAGEN公司DNeasyPlantMiniKits 5种方法提取基因组DNA ;并通过琼脂糖凝胶电泳、限制性内切酶处理和RAPD 3种方法对所提取的DNA样品进行检测 ,将它们在DNA的产量、质量和耗时、耗费等方面的优缺点进行定量总结 ,以便在实际工作中根据不同的实验目的选取最合适的方法 ,并根据分子生态学研究工作的实际特点确定了CTAB沉淀法为最佳方法。

Conifers are high in polysaccharides and other secondary metabolites such as polyphenols, terpens and resins, which makes it rather difficult to obtain high quality genomic DNA from their tissues. Five methods,i.e. basic method, high salt precipitation, CTAB precipitation, Ziegenhagen's method and DNeasy Plant Mini Kits(QIAGEN), were used to extract genomic DNA from the needle leaves of Pinus massoniana. The resulted DNA samples of the five methods were tested using agarose gel electrophoresis, restriction digests and PCR. The difference in yield, quality, time duration and expense of the obtained DNA are compared, and CTAB precipitation was decided as the best methed for the practical work of molecular ecology.