Russian Far East loaches of the genera Cobitis and Misgurnus are among members of the family Cobitidae with poorly understood systematics. In this study we present phylogenetic hypotheses based on mitochondrial (cytochrome b) and nuclear (RAG-1) sequences. All analyses recovered comparable topological phylogenies, and all data sets supported the non-monophyly of the genera Cobitis and Misgurnus. Both genera are represented by multiple lineages that in some cases do not correspond to the species described. We found some phylogenetic incongruities for the genus Misgurnus (M. mohoity and M. anguillicaudatus) that are explained by ancient hybridization, as was suggested previously for M. anguillicaudatus. The revealed phylogenetic relationships suggest that Paramisgurnus should be treated as a synonym of Misgurnus and M. bipartitus as a synonym of M. mohoity. All analyses recovered C. choii as a member of the genus Cobitis, confirming previous taxonomic conclusions. Most of the molecular lineages found follow currently recognized taxa with some exceptions, such as M. anguillicaudatus and C. lutheri. Phylogenetic relationships recover several unrelated lineages of M. anguillicaudatus and suggest additional studies to solve current taxonomic uncertainty. We found that C. lutheri is a non-natural group that contains two unrelated lineages: specimens of C. lutheri from the Far East of Russia collected close to the type locality and a second lineage with specimens of C. lutheri from Korea, the identification of which must be revised. The study provides evidence of the presence of the Far East species M. nikolskyi in Sakhalin Island, but simultaneously shows conspicuous genetic distinctiveness between the island and the mainland populations.