<em>Abstract</em>.—The systematics of lampreys was investigated using complete mitochondrial cytochrome <em>b</em> sequences from all genera and nearly all recognized species. The families Geotriidae and Petromyzontidae are monophyletic, but the family Mordaciidae was resolved as two divergent lineages at the base of the tree. Within Petromyzontidae, the nonparasitic <em>Lethenteron</em> sp. S and <em>Okkelbergia aepyptera</em> were recognized as distinct lineages, <em>Lethenteron morii</em> and <em>Lampetra zanandreai</em> were moved to new genera, a sister species relationship was recovered between <em>Caspiomyzon wagneri </em>and <em>Eudontomyzon hellenicus</em>, and a clade was recovered inclusive of <em>Entosphenus hubbsi</em> and western North American <em>Lampetra </em>(<em>L. ayresii </em>and <em>L. richardsoni</em>). The placement of <em>E. hellenicus</em> as the sister species to <em>C. wagneri </em>reduces the number of genera comprised entirely of parasitic species to two, <em>Geotria</em> and <em>Petromyzon</em>. The recognition of distinct lineages for <em>O. aepyptera</em> and <em>Lethenteron</em> sp. S recognizes, for the first time, lineages comprised entirely of nonparasitic species. Apart from the results mentioned above, monophyly was supported for the multispecific genera <em>Entosphenus</em>, <em>Eudontomyzon</em>, <em>Ichthyomyzon</em>, <em>Lampetra</em> (restricted to European species), and <em>Lethenteron</em>. Intergeneric relationships within Petromyzontidae were poorly resolved, but separate clades inclusive of <em>Entosphenus</em> and <em>Tetrapleurodon</em> (subfamily Entospheninae) and one comprised of <em>Eudontomyzon</em>, <em>Lampetra</em>, and <em>Okkelbergia</em> were recovered.