A large number of natural or synthetic dyes have been removed from both national and international lists of permitted food colors because of their mutagenic or carcinogenic activity. Therefore, this study aimed to use the Random Amplified Polymorphic DNA-Based Polymerase Chain Reaction (RAPD-PCR) assay as a feasible method to evaluate the ability of some food colors as genotoxin-induced DNA damage and mutations. Lactiplantibacillus plantarum was used as a bioindicator to determine the genotoxic effects by RAPD-PCR using M13 primer after treatment with some synthetic dyes currently used as food color additives, including Sunset Yellow, Carmoisine, and Tartrazine. Besides qualitative analysis, the bioinformatic GelJ software was used for cluster analysis to compare DNA fingerprints before and after treatment. The bacteria treated with the food colors showed the presence of polymorphism represented by DNA changes in the RAPD patterns, including variation in bands intensity, disappearance of normal bands, and appearance of new bands compared with the non-treated control. The GelJ program confirmed the presence of genetic variations between the bacteria treated with different concentrations of the food dyes and the bacteria without treatment. The RAPD approach can be applied for the detection of DNA damage and mutations induced by genotoxic compounds. Furthermore, L. plantarum and M13 are suitable as in vitro screening tools for detecting of potential genotoxicity of numerous compounds.