oa Dna Barcoding Of Lizards (reptilia, Squamata) Of Qatar

Recent rapid development in Qatar has brought awareness and recognition of the importance to catalogue, describe and preserve its biodiversity. Traditional methods of using morphological characteristics for species identification, although important, requires years of experience and is in need of complete specimens before identity is certain. Recent worldwide endeavor of DNA barcoding by using a segment of the mitochondrial gene cytochrome oxidase c 1 (CO1) to describe all species in the world, has gained recognition and popularity. In this student-centered research supported by Undergraduate Research Experience Program (UREP) of Qatar National Research Fund (QNRF), the objective is to sequence the barcoding gene CO1 of the lizards in Qatar and to compare the sequence among closely related species to facilitate future identification. The final results of this study will make available the barcode sequences of CO1 to be included in the GenBank database. Lizard species were collected across Qatar and photographs were taken for appropriated morphological identification. About 0.5 cm3 of muscle tissues were obtained from each specimen for DNA extraction. Currently between 8 to 12 different primer pairs suggested in literature and primer pair combinations were used for 7 species and a variety of programs of polymerase chain reaction (PCR) were employed in order to amplify the segment of gene of about 650 bp. Among the successfully amplified sequences from the 7 species currently analyzed, three species, Trachylepis septemtaeniata, Mesalina brevirostris, and Uromastyx aegyptia produced PCR products. To eliminate the presence of multiple sequences in the PCR product, PCR fragments were ligated to cloning vector in order to amplify one single sequence for each species. The current total success rate (<43%) based on the primer sequences obtained from publish results and from internal primers developed in this study further supports that the universal primers for reptiles are difficult to develop due to deep phylogenetic divergence. Future studies seeking universal primers should focus on candidate genes that evolve more slowly than CO1 gene to ensure higher homology of sequences among the reptilian species.