Xenarthra is a superorder of placental mammals constituted by extant armadillos, sloths, anteaters and their fossil relatives. Considering the biogeographic abundance, the two species included in the Tamandua genus are the most expressive among the anteaters, being found of the north of Central America to the south of South America. Tamandua tetradactyla stand out as the most expressive species of anteater, with often records in Brazilian biological collections. However, the absence of sexual dimorphism in the external morphology of T. tetradactyla and other anteaters contributes to that sexing mistakes are frequently observed in museum specimens. Thus, evolutionary, anatomical, ecological and pathological issues related to the sex of T. tetradactyla remain mostly unknown. Here, we investigate the use of PCR amplification of the SRY gene obtained from muscular tissues of T. tetradactyla in the sexual identification of museum specimens. In addition, we used the molecular data to reveal possible intraspecific sexual variations in the skeleton of the analyzed samples. The results indicate that the anatomical data are concordant with the molecular analysis in this study, with the appendicular bones of the specimens of T. tetradactyla presented marked patterns of sexual distinctions. The molecular-morphological integration is probably a useful tool in the recognition of intraspecific variation still unknown in museum specimens of Xenarthra.