| Summary: | This study was conducted at Head Panjnad, Pakistan to collect information about the available fish diversity. The evident fish orders found were siluriformes, anabantiformes, cypriniformes, cichliformes, osteoglossiformes, and synbranchiformes. The highest number of Hypophthalmicthys molitrix and the lowest number of Wallago attu were determined from this diversity. Simpson diversity index (D) and Shannon-Weiner index (H) were measured as 0.94938868 and 3.00940719, respectively. After the keen observation of various diversities at Head Panjnad, these fish species were selected for their COI gene and phylogenetic analysis. COI is the Cytochrome C Oxidase subunit 1 of mitrochondria (a gene sequence used in molecular investigations as a DNA barcode). The K2P (Kimura two-parameter) distances measured within species, genus, family, and order were 0.57%, 0.63%, 0.68%, and 0.77%, respectively. The K2P neighbor-joining tree was built on a commonly clustered species sequence in agreement with its taxonomic classification. The purpose was to create QR codes based on DNA sequences for accurate fish species identification. The current work concludes that COI sequencing might be used for fish species identification.
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