番茄SRAP-PCR反应体系建立与优化

Optimization for SRAP-PCR System of Tomato Based on Orthogonal Design

利用正交实验设计L16(45)对番茄SRAP-PCR反应体系的5个因素(Mg2+、dNTPs、引物、Taq DNA聚合酶和模板DNA)在4个水平上进行优化试验研究。结果表明:各因素水平变化对反应体系影响的大小依次为:Mg2+>dNTPs>引物>Taq DNA聚合酶>模板DNA;建立的番茄SRAP-PCR最佳体系(25μL)为:Mg2+2.5mmol/L、Taq DNA聚合酶0.5U、dNTPs0.25mmol/L、引物0.4μmol/L、模板DNA 80ng。

The SRAP-PCR system of Tomato was optimized by the orthogonal design which was in four levels of five factors（Mg2＋,Taq DNA polymerase,dNTPs,Primer,and DNA template）.The results showed that the order of each factor in different levels affected the result of PCR was：Mg2＋dNTPsprimerTaq DNA polymeraseDNA template;the optimum SRAP-PCR system（25 μL） for tomato included Mg2＋ 2.5 mmol/L,Taq DNA polymerase 0.5 U,dNTPs 0.25 mmol/L,Primer 0.4 μmol/L and DNA template 80 ng.