The first partial phylogeny of family Buthidae (17 genera) is presented, based on molecular data (16S rRNA mitochondrial DNA). The strong support for a monophyletic Old World group of 13 genera (mainly Palearctic desert forms) is demonstrated, while representative genera from Madagascar (Grosphus) and Southeast Asia (Lychas) group outside, as well as New World genera Centruroides and Rhopalurus. A very strong support is observed for the first time for three groups of Old World genera: (a) Compsobuthus, Mesobuthus, Liobuthus, Kraepelinia; (b) Hottentotta, Buthacus; (c) Orthochirus, Anomalobuthus. Phylogenetic hypotheses are discussed.