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¹ The Maharaja Sayajirao University of Baroda, Vadodara, Department of Zoology, Faculty of Science, - 390002 DOI : 10.4194/1303-2712-v17_5_22 Viewed : 6780 - Downloaded : 4507 The freshwater fishes of Gujarat have been largely untouched owing to the principal focus on the large marine sectors. In this paper, taxonomic identification has been done for few of the species and DNA barcoding has been attempted to strengthen the identification. This modern approach led us to the present study in which the classification of 52 species of the freshwater species found in selected six districts of Gujarat. 38 species of freshwater fishes, all belonging to the class Actinopterygii, were discriminated sequences (barcoded) for a 655bp region of the mitochondrial cytochrome oxidase subunit I gene (cox1). The samples were appropriately identified morphologically using standard available keys. Hence this will provide an insight of fish diversity and will take it to a further step, to carry out future molecular investigations which cannot be done if the sequences of the organisms are not known. Keywords : Taxonomy, freshwater fishes, COI gene