Background: Occupational exposures represent complex mixture of genotoxic agents. Benzene is one of the major organic product, people with chronic exposure to this organic product are exposed to many malignant disorders. People who work in Petrol station are consistently exposed to the dangerous components of petroleum through inhalation or direct contact with their skin. Objective: The aim of this study was to evaluate the potential cytogenetic damage and micronuclei frequency associated with occupational exposure to benzene among petrol station workers by using Micronucleus assay. Materials and Methods: The study groups included twenty five occupationally exposed petrol station workers and twenty five age matched controls were recruited and evaluated for genetic damage. Exfoliated Buccal cells were collected from both groups by using the sterile brush. Slides fixed in ethanol and stained with Giemsa stain and 1000 cells were studied by using a light microscope. Statistical analysis for data interpretation performed by Student’s t‑test, and P < 0.05 was considered statistically significant. Results: As compared to control subjects, significant high level of micronuclei frequency was found in the workers exposed to benzene (4.65±2 and 1.22±7 respectively). Period of exposure also showed considerable effects (P<0.05) on micronuclei frequency, while effect of age was non-significant (P>0.05). In this study, both Control and workers with history of Tobacco smoking and alcohol were excluded. Conclusions: The results of our study concluded that benzene have the potential to increase micronuclei frequency in the exposed subjects. The study also suggested that protective strategies should be implemented by the concerned authorities to minimize exposure to fuel.