Ning Setiati, FMIPA Biologi (2023) Polymerase Chain Reaction-Amplification of COI Gene of Stingray and Shark Using Four Primer Pairs. Tropical Journal of Natural Product Research, 7 (7). ISSN 2616-0684

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Abstract

The polymerase chain reaction (PCR)-based technique is essential for the successful amplification of the COI gene for the molecular identification of species. The present study was conducted to explore the amplification of the COI gene in stingray and shark samples using four primer pairs: FishF1-FishR1, FishF2-FishR2, FishF1-HCO2198R, and FishF2-HCO2198R. Five species of stingrays and two types of sharks were obtained from TPI Tasik Agung Rembang. DNA was isolated from the fish samples using the TIANamp Marine Animal DNA Kit. The concentration and quality of the DNA extracts were determined using the Nanodrop spectrophotometer and gel electrophoresis, respectively. The amplification results showed that the COI gene of Himantura gerrardi stingray was successfully amplified by the primer pair FishF1-FishR1; Himantura uarnocoides by the primer pairs FishF1-FishR1 and FishF2-FishR2; Himantura walga by the primer pairs FishF2-FishR2, FishF1-HCO2198R, and FishF2-HCO2198R; Neotrygon kuhlii by the primer pairs FishF1-FishR1, FishF2-FishR2, and FishF1-HCO2198R; Rhinobatos penggali by the primer pairs FishF1-FishR1, FishF2-FishR2, FishF1-HCO2198R, and FishF2-HCO2198R. Chiloscyllium punctatum and Carcharhinus sealei are shark species that were amplified successfully using the four primer pairs FishF1-FishR1, FishF2-FishR2, FishF1-HCO2198R, and FishF2-HCO2198R. The findings of this study reveal that the test primer pairs can be used for the molecular identification of stingrays and sharks.

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