Monophyly of the tribe Nycticeiini (Baeodon, Nycticeinops, Nycticeius, Otonycteris, Rhogeessa, Scoteinus [= Scoteanax and Scotorepens], Scotoecus, Scotomanes, and Scotophilus) has been challenged by new datasets over the last two decades including baculum morphology, cytogenetics, and mitochondrial ribosomal sequence data. These studies have resulted in new classifications for the Nycticeius-like bats, but only one study has empirically tested Nycticeiini monophyly. In this study, a suite of nuclear markers including both exon (APOB, DMPl, RAG2) and intron (PRKCI, STAT5A, THY) gene regions were used with previously studied mtDNA sequences (12S rRNA, tRNA^Val, 16S rRNA) to test Nycticeiini monophyly and develop new hypotheses for relationships of Nycticeius-like bats within Vespertilioninae. Although results of these phylogenetic analyses did not fully resolve phylogenetic relationships for all taxa historically included in Nycticeiini, they did reject the validity of Nycticeiini. Taxa historically circumscribed in this tribe were found scattered throughout generated phytogenies, with Scotoecus aligning basal to PipistrellusNyctalus, Nycticeinops with the Hypsugine group, Scotomanes with Eptesicus, and Rhogeessa with Antrozous.