The Optimization of the SSR Reaction System for Wild European Plum (Prunus domestica L.)

The research aims at obtaining the optimal SSR-PCR system for Wild European Plum, which is a kind of endangered fruit trees. The Wild European Plum was used as the research object, and an experimental design was employed to evaluate five factors (template DNA, dNTP, Mg2+, Taq DNA polymerase and primer) at six different levels. The results indicate that the optimal SSR-PCR reaction system for Wild European Plum was determined as follows: the optimal system of terminal volume 150 consisted of 0.4U Taq DNA polymerase, 0.4mM primers, 30-50ng tern-plate DNA, 0.3mM dNTP and 2.0mM Mg2+. The suitable thermal cycling conditions are melting at 94 degrees C for 5 min initially, followed by 35 cycles at 94 degrees C for 30 s, 50-59 degrees C for 45 s and 72 degrees C for 60 s, then keep the reaction mixture at 4 degrees C after a final extension step of 72 degrees C for 7-10 min. The optimized system would be effective as a solid foundation for Wild European Plum SSR analysis.