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DOI : 10.17693/yunusae.v17i26557.284131 Viewed : 3507 - Downloaded : 2225 Barbus grypus (Heckel, 1843) living in the Euphrates and Tigris river system, is one of the endemic fish species and has economic importance. In this study, genetic diversity of B. grypus populations was detected based on mitochondrial DNA cytochrome c oxidase subunit I (mtDNA COI) gene sequences. Three haplotypes and two polymorphic sites were detected in totally 36 individuals sampled from two populations. The average haplotype diversity (h) and nucleotide diversity (π) were calculated 0,246 and 0,00045 respectively. The result of Neutrality tests as statistically was insignificant (p>0.10). The results obtained by this study performed firstly for Barbus grypus species. Determined haplotypes for mtDNA COI gene region are new results for literature, in terms of genetic diversity of this species is an important data set. Keywords : Barbus grypus,Genetic diversity,mtDNA,Euphrates River,Tigris River