Background: DNA barcoding allows for species identification and description of genetic diversity. However, in the Middle East, information on genetic diversity is accumulating at a slower pace compared to that of other regions. Methods: The COI sequence of 24 lizard and snake species in Qatar that represent major families within the order Squamata were sampled and amplified via PCR using RepCOI primers (apart from one species). Purified amplicons were then aligned, and high- quality sequences were uploaded to BOLD. Using Sphenodon punctatus as the outgroup, the phylogenetic analysis was conducted using raxmlGUI software following the maximum likelihood method. Results: The COI sequence from each of the species was obtained and the consensus sequences were submitted to GenBank. In the phylogenetic analysis, a close relationship between members of the Agamidae and Serpentes was confirmed. While members of the same genus often showed sister-taxa relationships, and species in the same family were clustered with reasonably high bootstrap supports, the COI-based phylogeny was not able to resolve the relationships among genera within the families or identify relationships with high resolution at deeper lineages. Conclusion: Although ideal for species identification, COI gene sequencing is limited in phylogenetic inference due to high mutation rates that restrict its effectiveness for resolving relationships at deep phylogenetic levels. However, COI gene sequencing can be combined with nuclear markers for a more in-depth analysis.
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