Polychlorinated biphenyls (PCBs) are persistent pollutants in aquatic environments, often causing the decline or disappearance of wild populations. In this study, we used a random amplified polymorphic DNA (RAPD) assay to evaluate the effects on the genomic DNA of olive flounder embryo and larval stages of exposure to Aroclor 1254 at concentrations of 1, 5, 10, 20, and 40 μg/L. We compared RAPD fingerprints of exposed and non-exposed samples. Polymorphisms were revealed as the presence and/or absence of DNA fragments between the two samples. A dose-dependent increase in the number of polymorphic bands was observed with Aroclor 1254 treatment. Also, RAPD profiles of animals exposed to Aroclor 1254 exhibited an increase in the frequency values (FV) compared to the control. A phenogram constructed using neighbor-joining method indicated that genomic template stability in developing embryo and larval stages was significantly affected at ≥ 5 μg/L. This study suggested that DNA polymorphisms detected by RAPD analysis could be used as an investigative tool for environmental toxicology and as a useful biomarker in early life stages for the detection of potential genotoxicants.