Abstract: 【Objective】 In order to cater for the pressing needs of seed production units for high-throughput fast, accurate identification of the seed authenticity and purity, this research aims to establish an identification system for the hybrid watermelon cultivar Zaojia 8424 based on simple sequence repeat (SSR) markers. 【Methods】 Watermelon cultivar Zaojia 8424 and its two parents as research were taken as the testing materials, and polymorphic SSR markers characterized by distinct bands and stable amplification were screened, which would further optimize DNA extraction and PCR amplification systems. And also, a comparison between the aforementioned results and the results obtained in traditional field identification was performed to validate the accuracy of the proposed method. 【Results】 The four pairs of primers that generated complementary polymorphic bands for both parents were determined from 72 pairs of SSR markers on Zaojia 8424. The four pairs of primers were found to be distributed on the 5th, 6th and 9th chromosomes with segments ranging in size from 100 - 300 bp. The primer combinations with different amplified segments were conducted dual- and multiple- PCR amplifications, consequently, four PCR-amplified dual primer combinations were acquired. In addition, the SSR markers labeled as the combination WS27+MCPI-05 were used to identify the purity of 192 Zaojia 8424 plants grown in greenhouses, which well matched the result of traditional field identification. 【Conclusion】 The extraction of DNA using SSR markers and its combinations screened in this research integrated with an alkaline lysis method can be used for identification of purity of hybrid watermelon cultivar Zaojia 8424. Such method is proven efficient, and easy to operate, and applicable in a flow process of high-throughput screening, thus having a promising application potential.