海南安诺兰RAPD-PCR反应体系的建立

Establishment of Anota hainanensis RAPD-PCR Reaction System in Hainan

为海南安诺兰植物种质资源研究提供技术支持,以海南特有安诺兰种质为试验材料,采用正交试验,对影响RAPD分子标记的模板DNA、dNTPs、Taq聚合酶、Mg2+浓度和引物浓度5个因素进行优化,建立适合于安诺兰RAPD标记的反应体系。结果表明:5个因素优化后在20μL反应体积的浓度分别为模板DNA 60ng,dNTPs 150μmol/L,Taq酶1.0U,Mg2+2.0mmol/L,引物0.5μmol/L。

Five factors to influence the concentration of DNA template,dNTPs,Taq polymerase,Mg^2＋and primer of Anota hainanensis RAPD molecular marker were optimized by the orthogonal design to build the suitable reaction system of Anota hainanensis RAPD marker and to provide the technological support for research of Anota hainanensis germplasm resources.The results showed that the optimum concentration of DNA template,dNTPs,Taq polymerase,Mg^2＋and primer is 60 ng,150μmol/L,1.0U,2.0mmol/L and 0.5μmol/L separately.