Marine fisheries are important natural resources and must be maintained, especially fish species that are important sources of food. Despite the increase in stocking programs to maintain fisheries with artificially raised fish, the genetic impact stocking has on the wild fry population has not been addressed. Genetic variation in rock bream, Oplegnathus fasciatus, within and between wild-caught parents and the generation produced by them in 1 day was assayed using nine highly variable micro satellite markers. The nine micro satellite loci used in this study displayed diverse polymorphisms, and in total, 98 different alleles were observed over all loci. Differences in genetic variability of the offspring compared to their wild-caught parents (brood stock) were observed in terms of allele frequency, gene diversity, and heterozygosity. Although the generation of rock bream was missing 16% of the micro satellite alleles, no significant reduction was found in mean heterozygosity of the population compared to the brood stock. Eight of nine loci showed significant Hardy-Weinberg equilibrium (HWE) deviations in the population, while the brood stock deviated from HWE at three micro satellite loci (KOF85, KOF360 and KOF374). These deviations showed mostly a deficit of heterozygotes. Our results provide evidence for genetic differences in the hatchery offspring compared to their wild-caught parents and reinforce the need for a series of consecutive egg collections to avoid the loss of genetic variability. This also further underscores the importance of monitoring genetic variability of hatchery populations for the conservation of natural rock bream resources.