The present study aimed to evaluate the genetic relationships among the selected 12 freshwater fish species using the mitochondrial cytochrome C oxidase I (COI) gene sequences. The sequence data were retrieved from NCBI and analyzed the Multiple sequence alignment, Genetic distance estimation using the Kimura2-parameter model and phylogenetic tree construction using the neighbor-joining technique. The result revealed the conserved and the variable regions with in the COI gene and also showed tha