Tobacco is the most common misused drug in the world. In India, tobacco is used mainly in smokeless form. Tobacco is a potential carcinogen and its extensive use leads to generation of reactive oxygen species. Micronucleus assay has been used to assess the DNA damage. The aim of the present study was to set up the normal frequency of the presence of micronuclei and other nuclear anomalies in cells of the buccal mucosa and to compare it with tobacco chewers. Most of the tobacco chewers in the present study were mainly from rural areas, which indicates that the subjects living in rural areas are mainly indulged in the tobacco chewing habits. The number of normal cells was less in tobacco chewers and the number of nuclear anomalies was more, which clearly indicates the damage occurring due to tobacco chewing.