SSR标记小麦抗白粉病基因变性凝胶电泳体系的优化

为寻找适合SSR标记定位小麦抗白粉病基因的变性聚丙烯酰胺凝胶电泳的理想体系。主要研究Arc+Bis占凝胶的质量浓度、Arc/Bis质量比、上样量、电泳电压及染色等因素对变性聚丙烯酰胺凝胶电泳效果的影响。结果表明电泳效果较好的体系为:凝胶中Arc+Bis 60g/L、m(Arc)∶m(Bis)=29∶1、上样量5μL(含φ=20%的6×loading dye)、1 500V恒压、强碱法染色。此变性凝胶电泳体系下得到的图谱较为清晰。

An Optimal System of Denatured PAGE for Mapping A Powdery Mildew Resistance Gene in Wheat

To find an ideal system of denatured Polyacrylamide gel electrophoresis(PAGE)for mapping a powdery mildew resistance gene in wheat,the effect of total mass concentration of Arc and Bis in gel,mass ratio of Arc and Bis,sample volume,voltage and silver-staining methods on quality of electrophoretogram was studied.An optimal system of denatured PAGE was eventually established,which included 29∶1 mass ratio of Arc and Bis,60 g/L total mass concentration of Arc and Bis in gel,5 μL sample contained 20%(volume fraction) 6×loading dye,constant voltage 1 500V and silver-staining under strong alkali condition.The quality of electrophoretogram was improved while this system adopted.