DGGE技术检测中华绒螯蟹线粒体Cytb基因遗传多样性的实验条件探讨

目前，变性梯度凝胶电泳技术（DGGE）多用于微生物和临床医学检测，而用于水产动物基因片段检测的报道甚少。相关研究显示，针对不同的目的基因，其PCR条件、变性剂浓度梯度和电泳时间是DGGE检测实验成败的关键。因此，为应用DGGE技术分析中华绒螯蟹线粒体Cytb基因片段的遗传多样性，探讨了DGGE技术的关键因子，确定了PCR反应体系和循环参数、电泳时PCR产物的上样量、变性剂浓度范围和电泳的最适时间。

Study on Parameters of PCR and Electrophoresis in DGGE Technique on Analyzing Diversity of Mitochondrial Cytochrome b Gene of Eriocheir sinensis

The technology of Denatttring Gradient Gel Electrophoresis （DGGE） was used for microorganism and clinical medicine detection at present, the reports for detecting the gene fragments by DGGE technique in aquatic animals were extremely few. Some researches indicate that the PCR reaction parameters, the range of denaturant concentration of the gel and the times of electrophoresis are key factors in DGGE technique for analyzing different target gene fragments. In order to detect the diversity of mitochondrial cytochrome b gene of the Eriocheir sinensis using DGGE technique, some key factors, namely the suitable parameters of PCR reaction, the number of cycles, the loading quantities of PCR production, the range of denaturant concentration in polyacrylamide gel and the optimum electrophoresis time were confn-med.