The Dipsadidae contains more than 700 extant species belonging to 92 genera and is the largest family of American snakes. In this work, we built a data set including two mitochondrial genes (12S and 16S rRNA) for 125 dipsadid taxa belonging to 59 genera, in order to gain further insights on the phylogenetic relationships of this large group at the subfamilial and generic levels. Among dipsadines, the monotypic genus Nothopsis is the sister-group to Leptodeira. Among xenodontines, the monophyly of seven previously recognized tribes (Alsophiini, Elapomorphini, Hydropsini, Philodryadini, Pseudoboini, Tachymenini and Xenodontini) is confirmed. Among Xenodontini, the genus Liophis is paraphyletic with respect to Erythrolamprus and Umbrivaga and workers should be aware of the inadequacy of the current taxonomy. Finally, the following genera could not confidently be allocated to the above tribes: Caaeteboia, Echinantera and Taeniophallus, Tropidodryas, Manolepis and Pseudalsophis, Xenopholis, Psomophis, Hydrodynastes, Conophis and Crisantophis.