We developed 31 sets of melon (Cucumis melo L.) microsatellite markers in this study. Primers for these markers were designed from clones containing simple sequence repeats (SSRs) of (GA/CT)n or (GT/CA)n isolated from a melon genomic library. These microsatellite loci were examined for the detection of alleles among twelve lines and cultivars from six varieties of C. melo. A total of 28 marker loci showed polymorphism by 2.0 % agarose TBE gel separation. And these markers were also examined for cross-species amplification applicable to nine other species in Cucurbitaceae. Among them, 13 marker loci were commonly detected within the major cucurbit crops, cucumber, pumpkin and watermelon using the present primer sets. The species to which the melon microsatellite markers would be most applicable was Momordica charantia (24 markers), followed by Cucumis sativus (20 markers) and Cucurbita maxima (18 markers). These conserved microsatellite loci could be used as anchor markers in studies on synteny in Cucurbitaceae.