SSR标记小麦抗白粉病基因变性凝胶电泳体系的优化

摘要

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 60 g/L、m(Arc)∶m(Bis)=29∶1、上样量5 μL(含φ=20％的6×loading dye)、1 500 V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.%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 elec-trophoretogram 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 X loading dye, constant voltage 1 500V and silver-staining under strong alkali condition. The quality of electrophoretogram was improved while this system adopted.