小鼠胚胎胃肠道细菌基因组DNA提取方法比较

Comparative Study of Microbial Genome DNA Extraction Methods from the Gastrointestinal Tract of Murine Embryos

目的:筛选能均衡地提取小鼠胚胎胃肠道微生物区系各种细菌总DNA的方法。方法:分别采用反复冻融法、CTAB-SDS法、柱式基因组DNA提取试剂盒法提取小鼠胚胎胃肠道细菌基因组DNA,对其进行琼脂糖凝胶电泳、紫外分光光度计测定、PCR扩增等质量检测。结果:CTAB-SDS方法提取的基因组DNA纯度较高,OD260/OD280平均值最高,为1.845,电泳条带清晰,能满足下游的PCR扩增等分子操作。结论:确定CTAB-SDS方法为提取小鼠胚胎胃肠道细菌基因组DNA的最佳方法,为研究不同种动物胚胎的肠道菌群的结构和多样性奠定了基础。

Objective： To screen the method for balanced extracting microbial genomic DNA from the gastrointestinal tract of the murine embryos.Method： The repeated freeze thawing extraction method,hexadecyltrimethylammonium bromide-sodium dodecyl sulphate（CTAB-SDS） extraction method and TIANamp DNA mini kit extraction method were used to extract the genomic DNA from the gastrointestinal tract of the murine embryos.And DNA samples were evaluated through the agarose gelelectrophoresis,UV spectrophotometer analysis,PCR amplification respectively.Result：The methods based on the CTAB-SDS extraction buffer could obtain the higher purity and integrated gastrointestinal microbial genome DNA,the mean value of OD260/OD280 was 1.845.The electrophoresis strip of PCR amplification was clearing,and it could meet the need of downstream molecular applications.Conclusion： The CTAB-SDS method is preferred way for extracting microbial genomic DNA from the gastrointestinal tract of the murine embryos.This study would lay a foundation for further studies on the construction and diversity of intestinal microbial population from different mammalian embryo stage.