三种提取柱状黄杆菌基因组DNA方法的比较

采用酚氯仿抽提法、CTAB法和SDS-蛋白酶K法分别对鱼类病原菌柱状黄杆菌提取基因组DNA。使用超微量紫外分光光度计和琼脂糖凝胶电泳检测所提取的基因组DNA的产量和质量,并用PCR扩增对DNA进行了评价。结果显示,3种方法均可提取到柱状黄杆菌的基因组DNA,并能有效扩增细菌16S rDNA序列,但CTAB法提取的DNA产量和质量最高,CTAB法可以作为柱状黄杆菌DNA提取以开展分子生物学研究的首选方法。

Comparison of Three Methods for Extracting Genomic DNA from Flavobacterium columnare

The genomic DNA was isolated from fish pathogenic bacteria Flavobacterium columnare by phenol and chloroform method,CTAB method and SDS-protease K method.The quality and concentration of the genomic DNA were evaluated with ultramicrospectrophotometer,agarose gel electrophoresis and PCR amplification of the 16S rDNA.The result showed that genomic DNA can be obtained using the three different methods,and 16S rDNA were successfully amplified from samples extracted by the three methods.But the genomic DNA extracted with CTAB method had the best quality and yield.Therefore,the CTAB method was the best one to gain the genomic DNA of Flavobacterium columnare for the study of molecular level.