基于PCR DGGE的小鼠肠道菌群基因组提取方法的建立

通过比较4种小鼠粪便细菌总DNA提取方法对基于PCR-DGGE检测的肠道菌群多样性分析的影响，旨在建立适于PCR—DGGE的小鼠肠道微生物宏基因组提取的稳定、经济、快捷的方法。采用SDS裂解法、某国产市售粪便DNA提取试剂盒、改进的化学裂解法、改进的溶菌酶法4种方法提取小鼠粪便细菌总DNA，通过琼脂糖凝胶电泳、紫外分光光度法、细菌16S rRNAV3区PCR扩增结合DGGE对提取结果进行比较分析。SDS裂解法和国产市售试剂盒2种方法提取粪便细菌总DNA均未得到理想结果，另2种方法均能够检测到粪便中20种左右的细菌。改进的化学裂解法和改进的溶菌酶提取法的建立为基于PCR—DGGE进行肠道菌群结构的定量及定性分析提供了可靠的前提基础和实验保障。

Establishment of Method to Extract Intestinal Microflora Genomic DNA in Mice Based on PCR DGGE

Four methods to extract total mice fecal bacterial genomic DNA based on PCR-DGGE were compared to explore the influence of intestinal flora diversity analysis aiming at the establishment of a stable, economic and fast method suitable for intestinal microflora genomic DNA extraction in mice based on PCR-DGGE. SDS splitting method, some market-sold domestic fecal DNA extraction kits, advanced chemical splitting method, advanced lysozyme method were adopted to extract total mice fecal bacterial genomic DNA, the extraction results were analyzed comparatively through agarose gel electrophoresis, ultraviolet spectrophotometry, bacterial 16S rRNA V3 domain amplification combined with denatured gradient gel electrophoresis （DGGE）. No ideal extraction results were obtained adopting SDS splitting method and market-sold domestic fecal DNA extraction kits to extract mice total fecal DNA, however, another two methods could test about twenty bacteria from feces. Advanced chemical splitting method and advanced lysozyme extraction method provided premise basis and experiment assurance to establish quantitative and qualitative analyses of intestinal microflora composition based on PCR-DGGE.