果梅SSR反应体系的优化

Optimization on SSR analysis system of Japanese apricot( Prunus mume Sieb.et Zucc.)

以果梅品种小叶猪肝为试材 ,研究了果梅SSR技术中PCR反应体系的主要成分对SSR扩增结果的影响 ,并比较了采用聚丙稀酰胺凝胶及琼脂糖电泳检测扩增产物多态性的差异。结果表明 :在PCR反应体系中 ,Mg2 + 的最适浓度范围为1 89～ 2 83mmol·L-1;dNTP最适浓度为 0 2 4mmol·L-1;引物的最适浓度为 0 38μmol·L-1;Taq 聚合酶在 2 6 5 μL反应体系中宜加入 1U。利用此反应体系 ,对 2 4个果梅代表品种进行了SSR反应 ,用 8%的非变性聚丙稀酰胺凝胶电泳检测 ,扩增产物在 10 0～ 2 0 0bp之间 ,不同品种间DNA谱带多态性丰富。

The factors which affected on the SSR results of Japanese apricot and comparative analysis of polymorphism of SSR detected on two gel electrophoresis systems were studied.The results showed that the optimized content of Mg 2+ is 1 89?2 83 mmol·L -1 ;the optimized content of dNTP is 0 24 mmol·L -1 ;the optimized content of primer is 0 38 μmol·L -1 ;the optimized content of Taq polymerase is 1U·26 5 μL -1 in the PCR system.Through above PCR system,SSR fragments of 24 cultivars of Japanese apricot were obtained and detected.There were significantly different between two gel systems.Polyacrylamide gel(8%)had higher ability than agarose gel(3%)in discriminating amplified fragments.Polymorphism between different cultivars was abundant detected by Polyacrylamide gel.