Abstract: FengHua 3 was used in this study to optimize the several factors applied in SSR technique system. dNTP was found to have distinct effect to amplification, and annealing temperature specific to primer pair was also checked. Optimized SSR system for peanut was established : 25 mM MgCl2 1.5 μl, 2.5 mM dNTP 1.6 μl, 5 U/μl Taq 0.17 μl, 100 ng/μl template DNA 0.4 μl, 2.5 mΜ primers 5 μl were added to the PCR mixture. Optimized annealing temperature was 54℃.