Specific Primer Design for Detection of gene Cyt b for Shark Species Prionace glauca and gene COI for Carcharhinus spp. Using Real-Time PCR Method

Abstract

International market demand for sharks is increasing, thus increasing the number of catches of several shark species that are included in the International Union for Conservation of Nature’s (IUCN) Red List and the Convention International Trade of Endangered Species (CITES) Appendix II. Authentication using biomoleculars by utilizing DNA is necessary. This study was aimed to design specific primers based on cytochrome c oxidase I and cytochrome b marker genes for endangered sharks (Prionace glauca and Carcharhinus spp.) and to apply them in vitro to identify fishery products using real-time PCR techniques. This research begins with designing target species-specific primers. Designing a specific primer using BioEdit software. The Oligo Evaluator and NCBI’s Blast (Basic Local Alignment Search Tool) web tool for performing primary specification validation. The next stage is the sample preparation, DNA isolation, DNA amplification, DNA quality and quantity testing, and realtime PCR analysis. Primer’s design of target species Prionace glauca using cyt b gene and degenerate primer for genus Carcharhinus spp. COI gene markers were successfully carried out in silico. Efficient real-time PCR conditions of Prionace glauca and Carcharhinus spp. complies with testing standards using the real-time PCR method.