Molecular identification of Lutjanus species by PCR-RFLP analysis of mitochondrial 12S rRNA region

A PCR-RFLP was developed by targeting the mitochondrial 12S rRNA region to authenticate the six species viz., Lutjanus fulvus, L. rivulatus, L. quinquelineatus, L. fulviflanuna, L. madras and L. decussatus available along the Indian coast. A newly designed primer set, 12SU-F/12SU-R, was used for PCR amplification to obtain a product size of 550 bp, which was then digested using three endonucleases viz., BseDI, MnlI and MspAll. All the six species were unambiguously differentiated by BseDI enzyme. MnlI differentiated only four species viz., L fulvus, L. quinquelineatus, L. fulvifiamma and L. madras; while MspAI differentiated four species viz., L. rivulatus, L. quinquelineatus, L. fulviflamma and L. madras from the rest. The developed PCR-RFLP protocol using BseDI enzyme can therefore be easily adopted by regulatory authorities to the prevention of possible adulteration of species for red snappers.