Microsatellite is one of the most suitable markers for variety identification as it has great discrimination power for varieties with narrow genetic variation. The relationship between marker genotypes and 58 melon varieties was analyzed. Of the 400 pairs of simple sequence repeat (SSR) primers screened against 11 melon varieties. 28 pairs showed highly polymorphism on the basis of PIC (Polymorphism Information Contents) value. These markers were applied to constructing DNA profile data base of 58 major melon varieties through multiplex PCR and fluorescence based automatic detection system. A total of 111 polymorphic amplified fragments were obtained by using 28 SSR markers. The average of PIC value was 0.643 ranging from 0.401 to 0.846. One hundred eleven SSR loci were used to calculate Jaccard’s distance coefficients for UPGMA cluster analysis. A clustering group of varieties, based on the results of SSR analysis, were categorized into plant shape and fruit type. Almost all of the varieties were discriminated by marker genotypes. This information may be used to select comparative variety through comparison of genetic relationship between existing variety and candidate varieties in distinctive tests and protection of plant breeders’ intellectual property rights through variety identification.