We report the development of 11 microsatellite markers by an enrichment protocol in the ponerine ant Ectatomma tuberculatum. Polymorphism was explored in two colonies collected from Mexico and two colonies from Brazil. From the 11 loci which amplified, seven loci showed intracolonial polymorphism in Mexican colonies and only six loci were variable in populations from Brazil. Observed heterozygosity ranged from 0.18 to 0.84. The other five loci exhibited different alleles between Mexico and Brazil but geographical variability was not investigated further. Cross-amplification was tested in another species of the same Tribe (Gnamptogenys striatula) and one locus was revealed to be polymorphic.