药用植物雷公藤RAPD-PCR反应体系优化研究

Studies on Establishment of RAPD-PCR Reaction Conditions in Tripterygium wilfordii Hook.f.

采用单因子梯度试验法对影响雷公藤RAPD-PCR反应的Mg2+浓度、dNTPs浓度、引物浓度、Taq DNA聚合酶的用量及DNA模板浓度进行了筛选。结果表明获得清晰、重复性高的雷公藤RAPD-PCR扩增条件为:20μL PCR反应体积中,2.0 mmol·L-1 MgCl2,0.2 mmol·L-1 dNTPs,0.2μmol·L-1引物,50 ng模板DNA,1UTaq DNA聚合酶。

Some factors involving Mg^2＋, dNTPs, primer, Taq DNA polyrnerase and the templates were optimized for RAPD-PCR reaction system in Tripterygium wilfordii Hook.f with single-factor. The results showed that, in 20μL RAPD-PCR reaction system of Tripterygium wilfordii Hook.f, the most suitable concentration of MgCl2, dNTPs, template DNA, primer, Taq DNA polymerase were 2.0mmol·L^-1, 200μmol·L^-1, 50ng, 200nmol·L^-1 and 0.5U respectively.