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以苦瓜为试材,采用正交直观分析法和新复极差法相结合的方法,对影响苦瓜SRAP反应体系的5种因素(dNTP浓度、模板DNA、引物浓度、Mg2+浓度及Taq DNA聚合酶)4个水平进行优化筛选,以期优化苦瓜SRAP的PCR反应体系。结果表明:苦瓜SRAP分析的优化反应体系为20μL PCR反应体系中含有1×PCR buffer,250μmol/L dNTP,50ng模板DNA,1.2μmol/L引物,1.5mmol/L Mg2+,1.5UTaq DNA聚合酶。
Using bitter melon as the test material, five factors (dNTP concentration, template DNA, primer concentration, Mg2 + concentration and Taq DNA polymerase) that affect the bitter melon SRAP reaction system were determined by the method of orthogonality analysis and new repolarization method. 4 levels to optimize screening, in order to optimize the bitter melon SRAP PCR reaction system. The results showed that the optimum reaction system for SRAP analysis was 1 × PCR buffer, 250μmol / L dNTP, 50ng template DNA, 1.2μmol / L primer, 1.5mmol / L Mg2 + and 1.5UTaq DNA polymerase in 20μL PCR reaction system.