Background: Acorus calamus is a medicinal plant from India, which is used in the popular medicine for the treatment of malaria, arthritis, bronchial asthma, flatulent colic, and epilepsy. In this study, the genoprotective effect of A. calamus was evaluated in peripheral blood lymphocytes using cytokinesis block micronucleus assay (CBMN) and chromosome sensitivity analysis. Materials and Methods: Preliminary phyto chemical analysis of the extract was performed to determine its major phyto chemical constituents such as alkaloids, sterols, flavonoids, terpenoids, phenolic compounds, saponins, carbohydrates, glycosides, and tannins. Gas chromatography-mass spectrometry analysis of extract showed the presence of major bioactive compounds. Peripheral blood lymphocytes were treated with bleomycin (BLM) in the presence of methanol extract of A. calamus (MEAC) to observe its antigenotoxic potential. Results: In CBMN assay, treatment with MEAC at different concentrations in culture medium showed a statistically significant decrease in the frequency of micronucleus. A significant reduction in the chromosomal aberration frequency, especially chromosome gaps, breaks, acentric chromosomes, and dicentrics were also observed. Conclusion: The results clearly indicate that A. calamus can protect against DNA damage induced by BLM in cultured human peripheral blood lymphocytes and has proved to be a significant potential to protect cellular system from DNA damages.