Abstract: The orthogonal design was used to optimize SRAP-PCR amplification system on Clausena lansium on four levels of five factors (Taq polymerase,Mg²⁺,DNA template,dNTPs,and primer,respectively).The results indicated that the order of each factor in different levels affected on the result of PCR was Mg²⁺ or Taq polymerase,DNA template,primer and dNTPs.An optimal SRAP-PCR system was established,that was total 25 μl reaction system containing PCR10×Buffer 2.5 μl,0.75U DNA polymerase,2.0 mmol/L Mg²⁺,60 ng DNA template,0.2 mmol/L dNTPs,and 0.2 μmol/L primer.

Key words: Clausena lansium, SRAP, orthogonal design, optimization

摘要： 以黄皮(Clausena lansium)‘甜黄皮’品种为试材,利用正交设计L₁₆(4⁵)对黄皮SRAP-PCR反应体系中的5因素(Taq聚合酶、Mg²⁺、模板DNA、dNTPs、引物)在4个水平上进行优化试验。结果表明,不同因素对黄皮SRAP反应体系影响从大到小的顺序为:Mg²⁺和Taq聚合酶> 模板DNA> 引物> dNTPs;初步确立了适合黄皮的SRAP-PCR扩增体系为:在25 μl反应体系中,包括10×PCR buffer 2.5 μl、Taq DNA聚合酶0.75U、Mg²⁺ 2.0 mmol/L、模板DNA 60 ng、dNTPs 0.2 mmol/L、引物0.2 μmol/L。

关键词: 黄皮, SRAP, 正交设计, 优化