The Optimization of the SSR Reaction System for Wild European Plum (Prunus domestica L.)
THIRD CONFERENCE ON HORTICULTURE SCIENCE AND TECHNOLOGY (CHST 2012)(2012)
Abstract
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.
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Key words
Wild European Plum,SSR markers,Reaction system,Optimization
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