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DNA barcoding is a technique concerned with the classification of things that is done with the help of small gene or DNA sequence with a known location on a chromosome that can be used to identify individuals or species in organisms mitochondrial DNA (mt DNA) which is helpful in recognition of meticulous species. It uses sequence variety in a 658-base pair fragment near the 5? end of the mitochondrial cytochrome c oxidase subunit I (COI) gene as a means for species recognition. DNA barcoding is a more defined and steady method in comparison with the classification based on the form and structure of the organism. It is equally useful in any stage of life cycle of fishes. The present research project was designed to recognize tilapia fish species (Tilapia zillii) of Pakistan genetically. Blood samples (n=30) were collected from Tilapia fish and genomic DNA was extracted and confirmed by 1% agarose gel. A short segment of COI gene (680bp) was amplified. PCR products were sequenced and analyzed by bioinformatics tools. Number of the haplotypes was 7. The haplotype diversity was Hd: 0.584 while nucleotide diversity was Pi: 0.00244. The mean intraspecific K2P genetic distance was 0.019. The estimated transition/transversion bias R was 1.40 that showed that this species possess very low genetic diversity. COI may supply a landmark for the classification of associated species at molecular level. As tilapia is extensively used for food and many other purposes so use of DNA barcoding technique is very helpful in discriminating it from other correlated fish species. It will also reduce the chance of mislabeling of tilapia fish species during its trading internationally as well as the species assessment at national level.
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