一种快速提取肠道微生物总DNA的方法

A rapid method for total DNA extraction from microorganisms of intestinal tract

采集的兔肠道内容物及其粪便样品,通过分散浸泡、震荡洗涤、分级离心、滤器过滤、DNA提取试剂盒提取纯化,可以获得纯度很高的DNA样品。经0.8%琼脂糖凝胶电泳检测和紫外分光光度计测定,样品A260/A280的比值为1.72±0.02。分别以提取的DNA样品为模板,通过设计的细菌特异引物,对其16S rDNA基因进行PCR扩增,获得了1.6 kb大小特异性很好的预期条带。这为肠道微生物群落的分子生态学研究提供了一种简便、可靠的DNA提取方法。

Some faecal samples were collected from intestinal tracts of rabbits. These samples were soaked, vibrated and washed in the specific buffer solution. After graduation centrifuged, the supematant was filtered with G3 glass tundish and extracted with Bacterial DNA Kit, and the high-quality total DNA from a wide variety of bacterial species was isolated and determined by a spectrophotometer （A260/A280 = 1.72±0.02） and 0. 8% agarose gel electrophoresis. The purified DNA was suitable for PCR, restriction digestion and hybridization techniques. The 16S rDNA was amplified from these DNA samples through a set of bacteria-specific primers. The molecule of PCR products was about 1.6 kb. This protocol had potential for molecular microbial ecology work in gastrointestinal tract.