美花石斛SSR-PCR体系的优化研究

Optimization of SSR-PCR System of D.loddigesii Rolfe

采用正交设计法对影响美花石斛SSR-PCR体系的5个主要因素进行了优化,建立适宜的美花石斛SSR-PCR的反应体系,并利用黔产5个不同的美花石斛居群共计48株进行了验证。结果表明:在10μL的美花石斛的反应体系中各因素的最佳浓度分别为2.0mmol/L Mg^(2+),0.4mmol/L dNTP,1.5UTaq酶,0.4μmol/L引物,40ng模板DNA,建立的SSR-PCR体系可用于美花石斛的遗传多样性研究。

In order to establish the best SSR-PCR reaction system, by the method of the orthogonal design affect on D. loddigesii SSR-PCR system of 5 elements（Mg2＋ , dNTP, Taq enzyme, primers and DNA） was optimized,which estabiished suitable D.loddigesii SSR-PCR reaction system.And a total of 48 species of D. loddigesii Rolfe from 5 group in guizhou were tested.The results showed the optimum concentration of each factor in the reaction system of D.loddigesii in I0 pL volumes was 2.0 mmol/L Mg2＋ ,0.4 mmol/L dNTP, 1.5 U Taq DNA polymerase,0.4μmol/L primer and 40 ng template DNA.The established SSR-PCR system can be used for analyzing genetic diversity of D,loddigesii.