蛋黄果ISSR-PCR反应最佳体系的构建(简报)

Optimization of ISSR-PCR System for Lucuma nervosa by Using Orthogonal Design

利用正交设计的方法,对蛋黄果ISSR-PCR反应的5因素4水平进行试验,试验结果运用SAS9.0软件进行分析。结果表明,在20μL的反应体系中,蛋黄果ISSR-PCR反应的最佳体系为:Taq酶0.75U,dNTPs0.2mmol/L,模板80ng,引物0.25μmol/L,Mg2+1.5mmol/L。

An orthogonal design was arranged to optimize ISSR-PCR amplification for Lucuma nervosa A. DC at 4 levels of five factors. The results of PCR were analyzed by software SAS9.0, and the analysis showed that the most suitable ISSR-PCR system for Lucuma nervosa A. DC. contained 0.75 U Taq DNA polymerase, 0.2 mmo1/L dNTP, 80 ng DNA template, 0.25 μmol/L Primer and 1.5 mmo1/L Mg2+ in a 20 μL reaction substrate.