鹅肠道微生物区系的PCR-SSCP体系构建  

Establishment of single-strand-conformation polymorphism(SSCP) for microbial community Structure in goose intestine

作　　者：徐敏娟[1] 刘蓓一[1] 王志跃[1] 周秋燕[1] 范莉[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《草业科学》2008年第12期99-103,共5页Pratacultural Science

摘　　要：聚合酶链反应-单链构象多态性分析(PCR-SSCP)技术是利用DNA单链构象具有多态性进行基因检测的一种快速、简便、较灵敏的分析技术,但其影响因素很多。研究运用PCR-SSCP技术分析鹅肠道微生物区系,探讨了凝胶浓度,胶联度,PCR产物的稀释倍数,电泳时间和温度,是否加甘油、尿素和λ-核酸外切酶等因素对SSCP图谱的影响。结果表明,凝胶浓度为12%,胶联度为39∶1,凝胶中不加甘油,电泳电压为120 V,在恒温环境中,试验重复性较好,条带较清晰。因而,在采用PCR-SSCP技术分析不同的研究对象,需要对其参数进行最大程度的优化,才能获得试验的成功。PCR-SSCP is one of rapid, simple and sensitive analytic technology to examine genome adopting DNA single strand conformation. However, many factors influence the results of SSCP analysis. In order to study microbial community in goose intestine, various controlled conditions such as the gel-concentration, the concentration of cross-linker, the dilution of the PCR products, the electrophoresis temperature, voltage, the presence of glycerol, urea and Lambda exonuclease in gels during elec- trophoresis were studied. Results showed that while the gel-concentration is 12% （acrylamide-bisac- rylamide 39： 1）, no glycerol, and the voltage is 120 V, with constant temperature during electrophoresis were the best SSCP conditions.

关 键 词：鹅 微生物 PCR-SSCP 

分 类 号：S852.6[农业科学—基础兽医学]