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油菜SSR检测是其主要农艺性状分子标记及标记辅助育种的一项重要技术。以无花瓣品系APL0 1、常规有花瓣抗菌核病品种M 0 83及其回交群体BC1为供试材料 ,研究油菜SSR检测体系中PCR扩增的各主要成分对检测结果的影响 ,比较不同电泳方法及染色方法的效果 ,进一步优化了适用于油菜的SSR检测体系。优化后的PCR反应体系为 :1×Buffer(含 2 0 0mmol/LMgCl2 )、5 0 0 0 μmol/LdNTPs、0 5 0UTaq酶、0 0 8μmol/LSSR引物、6 0 0ng/μl模板DNA ,加ddH2 O至 10 0 0 μl。对PCR产物的电泳染色采用聚丙烯酰胺凝胶电泳及银染
SSR detection of rapeseed is an important technique of molecular marker and marker-assisted breeding of its main agronomic traits. The effects of the main components of PCR amplification in rapeseed SSR detection system on the detection results were compared with the APL0 1 without petal line, the conventional petal disease resistant line M 0 83 and the BC1 backcross population, Methods and staining methods to further optimize the application of rapeseed SSR detection system. The optimized PCR reaction system consisted of 1 × Buffer (containing 200 mmol / L MgCl 2), 500 μmol / L dNTPs, 0 05 Ohq enzyme, 0 0 8 μmol / LSSR primer and 600 ng / μl template DNA plus ddH 2 O to 100 μl. The electrophoresis of PCR products using polyacrylamide gel electrophoresis and silver staining