Seventeen new polymorphic microsatellites were isolated and characterized for the South American Rufous-crowned Elaenia, (Elaenia ruficeps). E. ruficeps is a bird commonly found in white sand vegetation, a naturally fragmented and threatened Amazonian habitat. The number of alleles per locus ranged from 2 to 20, whilst the observed and expected heterozygosities varied from 0.074 to 0.967 and from 0.173 to 0.919, respectively. Seven loci significantly deviated from Hardy–Weinberg equilibrium. Eleven microsatellite loci were cross-amplified in at least one additional Tyrannidae species. These microsatellite markers will be useful tools for further studies of avian population structure and genetic variability in this unique habitat.