The co-amplification of conserved numts mask the divergent mitochondrial D- loop gene

Background and Aim: Mitochondrial DNA has been used as the preferred genome in biodiversity research. However, several factors contribute to its inadequacy. The occurrence of nuclear pseudogenes known as numts. Numts establish one of the main difficulties that prevent obtaining orthologous mitochondrial sequences. The current study aimed to test the efficiency of standard PCR protocol in producing orthologous sequences of control region in Mellivora capensis, a carnivore mammal inhabiting the southern part of Iran.Methods: Tissue samples were collected, across its distribution range in Iran. DNA extractions were performed in the laboratory using Genomic DNA Mini Kit. Extracted DNA from each sample was used to amplify two different fragments of D-loop region (۲۵۰ and ۵۸۰ bp) by standard PCR protocol.Results: The PCR amplicons were checked for numt possibility. Results showed the existence of numts in D-loop gen in this species. The first evidence of the mtDNA numt was the existence of non-specific PCR amplification. The reactions using primers designed to amplify mtDNA copies produced variable sequences among individuals. For the first (۲۵۰ bp) fragment, numt signs were observed in all amplicon, however, the numt percentage decreased for the longer fragment.Conclusion: Due to little information available on the species, it is required to conduct more studies on the species, using large sample size to better identify genetic diversity of the species. This paper suggested that Microsatellite method to find genetic diversity in Mellivora capensis.

کلیدواژه ها:

Mitochondrial DNA ، Numt ، D-loop ، Mellivora capensis