黄瓜SSR-PCR反应体系的优化

Optimization of SSR-PCR reaction system in cucumber

采用L9(34)正交试验设计,研究了黄瓜SSR-PCR反应体系的主要成分对扩增结果的影响,并比较了变性聚丙烯酰胺凝胶电泳和琼脂糖凝胶电泳检测扩增产物多态性的差异。结果表明,最适反应体系为:在20μL体系中包括dNTP 200μmol.L-1、Primer 0.4μmol.L-1、Mg2+2.5μmol.L-1、Taq酶0.5μmol和模板DNA60 ng。用变性聚丙烯酰胺凝胶电泳检测SSR扩增产物结果更准确。

The factors which affected on the SSR-PCR results of cucumber were studied with L9 （34） orthogonal design. And comparative analysis of polymorphism of SSR detected on denaturing polyacrylamide gel electrophoresis and agarose gel electrophoresis. The result showed that the optimal reaction system was as follows： the 20 μL reaction system contained dNTP 200 μmol.L^-1, Primer 0.4 μmol.L^-1, Mg2. 2.5 μmol.L^-1, Taq DNA polymerase 0.5 μmol.L^-1 and DNA template 60 ng. The detection effects of denaturing polyacrylamide gel electrophoresis were more accurate.