Abstract: Polydomy occurs when an ant colony occupies at least two spatially separated but socially connected nests. The identification of polydomy in ant colonies is important to understand ant life history, evolutionary success, and biotic and abiotic impacts on the environment. Several methodologies are used for the correct delimitation of the borders of the colonies and, therefore, in the identification of colonies with polidomous structure, these limits can be determined according to behavioral, spatial and genetic approaches. In the present study we develop and characterize microsatellite markers in order to complement ecological results that suggest the existence of polydomy in the cerrado ant species, Camponotus leydigi Ver menos