Polymerase Chain Reaction-Amplification of COI Gene of Stingray and Shark Using Four Primer Pairs: http://www.doi.org/10.26538/tjnpr/v7i7.3

Ning Setiati; Ning Setiati; Ning Setiati; Partaya Partaya; Nur Hidayah; Priyantini Widiyaningrum

Authors

Ning Setiati Department of Biology, Universitas Negeri Semarang, Semarang 50229, Central Java, Indonesia
Ning Setiati Department of Biology, Universitas Negeri Semarang, Semarang 50229, Central Java, Indonesia
Ning Setiati Department of Biology, Universitas Negeri Semarang, Semarang 50229, Central Java, Indonesia
Partaya Partaya Department of Biology, Universitas Negeri Semarang, Semarang 50229, Central Java, Indonesia
Nur Hidayah Department of Biology, Universitas Negeri Semarang, Semarang 50229, Central Java, Indonesia
Priyantini Widiyaningrum Department of Biology, Universitas Negeri Semarang, Semarang 50229, Central Java, Indonesia

Keywords:

Stingray, Shark, Primer, PCR, COI gene, Amplification

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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