Abstract: The study took strawberry variety‘Fengxiang’as tested material and used one-factor experimental design to establish the SRAP-PCR reaction system of strawberry, which including four different gradients of Mg²⁺ , dNTPs, TaqDNA polymerase, primer concentration, and based on this, optimized the template DNA concentration and annealing temperature. The results showed that the optimum SRAP- PCR system of strawberry was as follows: 20μL capacity of reaction system contained 10×PCR buffer 2μL, Mg²⁺ 2.0 mmol/L, dNTPs 0.3 mmol/L, forward primer and reverse primer 0.6μmol/L, Taq DNA polymerase 1.0 U, template DNA 100 ng. The SRAP- PCR amplification procedure was as follows: pre- denaturation at 94℃for 5 min; denaturation at 94℃for 1 min, anneal at 35℃for 1 min, extension at 72℃for 1 min and in total 5 cycles; denaturation at 94℃for 1 min, anneal at 54℃for 1 min, extension at 72℃for 1 min and in total 35 cycles; extension at 72℃for 5 min; preservation at 4℃. Twenty-nine primer pairs were screened out from 110 SRAP primer pair combinations based on their clear and rich amplification bands and good polymorphism by utilizing the optimal SRAP reaction system. So the optimal system was reliable which could be applied in the germplasm identification and molecular marker assisted breeding of strawberry.