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本实验以产于四川、安徽、广东和青海的白芥农家品种为材料,采用单因素筛选和L16(45)正交设计相结合的方法,对白芥ISSR-PCR反应体系的Mg2+浓度、dNTPs浓度、引物浓度、Taq酶和模板DNA用量进行优化,建立最佳的白芥ISSR-PCR反应体系.结果表明:在20 μL的PCR反应体系中,Mg2+浓度2.0 mmol/L、dNTPs浓度0.5 mmol/L、TaqDNA聚合酶0.2 U、引物浓度0.4 μmol/L、模板DNA 60 ng为最佳用量.用4个白芥农家品种对建立的ISSR-PCR反应体系进行验证,表明该反应体系稳定性高、可重复性好,同时筛选出条带清晰、多态性较好的18条引物.该反应体系的建立为白芥ISSR标记开发、种质资源鉴定与筛选、遗传多样性分析和分子标记辅助育种等提供了帮助.“,”In this study,Sinapsis alba from Sichuan,Anhui,Guangdong and Qinghai were selected as materials.We applied the combination of single factor experiment and L16(45) orthogonal experiment to optimize the concentration of Mg2+,dNTPs,primer,content of Taq polymerase and concentration of template DNA in the ISSR-PCR reaction system of Sinapsis alba,aiming to establish the best ISSR-PCR reaction system of Sinapsis alba.Results showed that the optimized 20 μL ISSR-PCR system included Mg2+ 2.0 mmol/L,dNTPs 0.5 mmol/L,Taq polymerase 0.2 U,primer 0.4 μmol/L and template DNA 60 ng.The optimized system was then verified by four different Sinapsis alba varieties,the results of which indicated that the system was stable and reproducible.Meanwhile,18 primers with clear bands and good polymorphism were screened.The establishment of this reaction system offer help for the development of ISSR markers,identification and selection of genetic resources,genetic diversity analysis and molecular marker assisted breeding ofSinapsis alba.