鹌鹑肠道微生物总DNA的最佳提取方法

The best method for the extraction of total DNA from the intestinal microbes in quails

为了获取高质量、较完整的肠道菌群基因组DNA,试验采用常规的饱和苯酚/氯仿法(方法一)、牛瘤胃DNA的提取法(方法二)和对上述两种方法进行优化后得到的肠道微生物DNA的提取法(方法三)对鹌鹑肠道微生物总DNA进行了提取,并对结果进行了比较。结果表明:应用方法一和方法二提取的DNA浓度比较低,电泳显示样品DNA条带没有方法三明亮;以方法三提取的肠道总DNA为模板,采用细菌通用引物,对其16S rDNA V3可变区进行PCR扩增,得到较清晰的图谱,条带整齐。说明采用方法三提取鹌鹑肠道微生物DNA较完整,可用于后续的分子生物学试验研究。

To obtain high -quality and relatively complete genomic DNA of the intestinal flora , in this study,the experiment used three methods to extract the DNA. The first was a conventional method for using saturated phenol / chloroform, the second was a method in accordance with the DNA extraction method from rumen of cattle, and the third was a method for optimizing the first two methods to explore the best extraction method of total DNA of the intestinal flora in quails. The results showed that the concentrations of DNA extracted with first two methods were lower than that of the third method. The electrophoresis results showed the I）NA bands were not as bright as that of using the third method. Tak- ing the total DNA extracted from the intestine as a template using the third method, the 16S rDNA was amplified from these DNA samples through a set of the bacteria - specific primer, which obtained the clear atlas and neatly bands. It indicates that the DNA extracted from the in- testinal microbes in quails is relatively complete using the third method, and the method can be used for molecular biology research.