应用DGGE研究微生物群落时的常见问题分析

Common problems in the analyses of microbial community by denaturing gradient gel electrophoresis(DGGE)

变性梯度凝胶电泳(DGGE)是通过核酸片段对微生物群落进行研究,可以监测未培养细菌及其功能基因,被广泛地应用于微生物群落多样性和动态分析,并成为微生物分子生态学研究中的重要手段之一。文中论述了DGGE操作过程中遇到的常见问题,并提出了相应的解决方法。全面分析了样品预处理过程和PCR扩增效果对DGGE分析的影响,探讨了DGGE图谱的优化过程和图谱分析方法,并对DGGE的应用前景进行了综述。

Denaturing gradient gel etectrophoresis （DGGE） can describe microbial communities based on partial sequences of nucleic acid and monitor uncultured bacterium and it＇ s functional genes. It is well-established molecular tools in microbial molecular ecology that allows the study of diversity and dynamics of microbial communities, Here, common issues in the analyses of microbial community by DGGE were raised, and the feasible resolving methods were introduced in this paper. The effects of samples pretreatment and PCR amplification on DGGE analysis were evaluated, Optimization and statistical analysis of DGGE profiles were discussed. Finally, prospect of DGGE for understanding microbial communities were reviewed.