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Not AvailableDNA barcoding has been widely used in species identification and biodiversity research because it has been shown that in many groups, including insects, interspecific variation in DNA sequences of some genes is much higher than intraspecific and this provided an opportunity to use DNA sequences for species identification. Cytochrome oxidase I (COI) barcoding sequences can be used to discover cryptic, closely related and morphologically similar species. DNA barcoding has gained increased recognition as a molecular tool for species identification in various groups of organisms. A study was, therefore, undertaken to barcode five fly species prevalent in poultry farms in and around Bengaluru districts in Karnataka state. The barcoding of COI gene of Musca domestica, Chrysomya megacephala, Hydrotaea capensis, Hermetia illucens and Sarcophaga ruficornis yielded an amplified fragment of 658 bp sequence. Barcode for all 5 species was generated using Bold_Systems v3 and submitted to GenBank and accession numbers were obtained. In the present study, identification of five different fly species based on morphology was also confirmed by DNA barcoding to prove their correct identity.Not Availabl