Pseudophoxinus firati Bogutskaya, Kucuk & Atalay, 2007 (Actinopterygii, Cyprinidae) living in the Euphrates River system were analyzed in terms of their karyotype, C-banding, and nucleolus organizer region properties. Metaphase chromosome spreads were obtained from kidney cells. The diploid chromosome number was found to be 2n = 50, of which 19 pairs were meta-submetacentric and 6 pairs were subtelocentric, and the fundamental number was found to be 88. No sex chromosome differentiation was observed. C-banding revealed heterochromatin in the pericentromeric regions of 6 pairs of chromosomes. NORs were identified on the short arms of 2 pairs of medium-sized submeta-subtelocentric chromosomes. This study may contribute to other cytogenetic research.