Strathdee Fiona, Free Andrew
Institute of Cell Biology, University of Edinburgh, Edinburgh, UK.
Methods Mol Biol. 2013;1054:145-57. doi: 10.1007/978-1-62703-565-1_9.
Denaturing Gradient Gel Electrophoresis (DGGE) is a technique used to separate short- to medium-length DNA fragments based on their melting characteristics. It has been used frequently for identifying single-nucleotide polymorphisms without the need for DNA sequencing and as a molecular fingerprinting method for complex ecosystem communities, in particular in conjunction with amplification of microbial 16S rRNA genes. Here, the principles of DGGE, based on partial DNA strand separation at a given position in a gradient of chemical denaturant, are described, and an example protocol, optimized for fingerprinting of 200-300 bp fragments of bacterial 16S rRNA genes, is given.
变性梯度凝胶电泳(DGGE)是一种用于根据短至中等长度DNA片段的解链特性对其进行分离的技术。它经常被用于无需DNA测序即可鉴定单核苷酸多态性,并且作为一种用于复杂生态系统群落的分子指纹识别方法,特别是与微生物16S rRNA基因扩增相结合时。本文描述了基于在化学变性剂梯度中给定位置的部分DNA链分离的DGGE原理,并给出了一个针对细菌16S rRNA基因200 - 300 bp片段指纹识别进行优化的示例方案。