【摘要】 应用L16(45)正交设计对影响马铃薯SSR-PCR的主要参数进行优化,建立适于马铃薯SSR-PCR反应的最佳体系。结果表明:各因素不同水平浓度对PCR反应结果均有显著影响。马铃薯SSR-PCR优化反应体系为:10×PCR Buffer 1.0μl、模板DNA约30 ng、dNTP 200 mol/L、SSR引物66 ng、Taq DNA聚合酶0.25 U、Mg2+2.25 mmol/L,加ddH2O至终体积10.0μl。PCR扩增程序如下:94℃预变性5 min;94℃变性1 min,53℃复性1 min,72℃延伸1.5 min,共35个循环;72℃延伸10 min,然后4℃保存。更多还原

【Abstract】 Used L16(45) orthogonal designs for optimizing the main factors of the SSR-PCR reaction system of potato and establishing the optimum SSR reaction system finally.The result showed that there were significant effects on the results of SSR-PCR at different concentration of each factor,and the optimized SSR-PCR reaction system was:10×PCR Buffer 1.0μl,DNA template 30 ng,dNTP 200 μmol/L,SSR primer 66 ng,Taq DNA polymerase 0.25 U,Mg2+ 2.25 mmol/L.,adding ddH2O to terminal volume 10.0 μl.The PCR amplification procedure was:predenaturation for 5 min at 94 ℃,followed by 35 cycles of denaturation for 1 min at 94 ℃,anneal for 1 min at 53 ℃,extension for 1.5 min at 72 ℃,the amplification was completed after extension for 10 min at 72 ℃,then stored at 4 ℃.更多还原