濒危植物海南龙血树SSR-PCR反应体系优化

Optimization of SSR-PCR Reaction System in Dracaena cambodiana,An Endangered Plant

为了更好地应用SSR分子标记技术于濒危植物海南龙血树保护遗传学研究,本研究通过单因子试验与正交试验方法,优化建立海南龙血树SSR-PCR反应体系,并对优化后体系的稳定性进行了检测。研究结果表明,优化后的海南龙血树15μL SSR-PCR反应体系为:1.5μL 10×PCR buffer, Mg^(2+)浓度为2.0 mmol/L,d NTPs浓度为100μmol/L,Taq酶1.0 U,引物0.4μmol/L和DNA模板5 ng。经验证,该反应条件可用于海南龙血树遗传多样性和遗传结构分析。

In order to better use the SSR molecular marker in the conservation genetics study of Dracaena cambodiana,this study used single factor test and orthogonal test method to optimize SSR-PCR reaction system in D.cambodiana,and the stability of the optimized system was tested.The results showed that the optimized 15 μL SSR-PCR reaction system of D.cambodiana included 10×buffer 1.5 μL,2.0 mmol/L MgCl2,100 μmol/L d NTPs,Taq DNA polymerase 1.0 U,Primer 0.4 μmol/L and DNA template 5 ng.After verification,the optimized SSR-PCR reaction system was suitable for the analysis of genetic diversity and genetic structure of D.cambodiana population.