杏SSR反应体系的优化研究

Optimization of SSR System in Apricot

研究了杏SSR-PCR反应体系的主要成分对扩增结果的影响,并进行了体系验证。优化后的反应体系为：总体积20μL,1×buffer2、.0 mM/L Mg^2＋0、.25 mM/L dNTP、0.20μM/LPrimer6、0 ng/20μL模板DNA和0.05 U/μL Taq酶。利用32个杏品种验证此反应体系,6%的变性聚丙稀酰胺凝胶电泳检测结果显示,扩增产物在184-267 bp之间,不同品种间DNA谱带具有多态性,且反应体系的稳定性和可重复性好。

The factors which affected the SSR--PCR results of Apricot were studied in the experiment. The reaction system optimized was as follows ： the 20 μL reaction system contained Mg^2＋ 2. 0 mM/L, dNTP 0.25 mM/L, Primer 0.20 μM/L,DNA tamplate 60 ng/20μL,Taq DNA polymerase 0. 05 U/μL and PCR buffer 1×. To evaluate this SSR system, 32 accessions of apricot were used. The 6 percent polyaerylamide gelelectrophoresis showed that the PCR products were between 184-267 bp and polymorphic in DNA among the apricot accessions, indicating that the SSR reaction system was steady and reproducible.