Application of mitochondrial gene markers and real-time PCR for the identification of Isurus spp. and Lamna nasus species

Department of Aquatic Product Technology, Faculty of Fisheries and Marine Sciences, IPB University, Bogor 16680, Indonesia

^* Corresponding author: asabdullah@apps.ipb.ac.id

Abstract

Sharks are predator species that play an important role in marine ecosystems and have high economic value. Exploiting shark parts, including fins, scales, teeth, cartilage, skin, liver, meat, and tails, is difficult to identify visually, necessitating molecular identification to determine traded shark products. This study aimed to design species-specific primers based on mitochondrial gene markers for shark species Isurus spp. and Lamna nasus, and their application in real-time PCR-based fisheries product authentication. The research was conducted in silico for specific primer design and in vitro for DNA sample isolation, DNA quantity and quality testing, DNA amplification, and real-time PCR analysis. The primer pairs of species included in the CITES II list with different genetic markers have been successfully created. The primers used were able to amplify DNA and showed a Ct difference between the target and non-target DNA, proving their ability to identify the target shark species.