PCR-RFLP Analysis of 12s and 16s Mitochondrial rRNA Genes from Brackishwater Finfish and Shellfish Species

Abstract

The use of traditional genetic characterization techniques for detection of genetic variation in aquatic species based on morphological characters has its own limitations. One of the modern approaches to study genetic variation in aquatic species is by Restriction Fragment Length Polymorphism (RFLP) of mitochondrial DNA (mtDNA). Amplification of mitochondrial 12s and 16s rRNA genes from tiger shrimp (Penaeus monodon, Penaeidae), white shrimp (Fenneropenaeus indicus, Penaeidae), grey mullet (Mugil cephalus, Mugilidae), tilapia (Oreochromis mossambicus, Cichlidae), Asian sea bass (Lates calcarifer, Centropomidae) and mud crabs (Scylla serrata and Scylla tranquebarica, Portunidae) and the characterization of the amplified PCR products by RFLP have been carried out in the present study. The use of the primers in the present study was found to be universal for amplification of mitochondrial 12s and 16s rRNA genes across the taxonomically different brackishwater species examined in this investigation. The amplified products of 12s and 16s rRNA mitochondrial gene segments obtained with these primers can be used for restriction digestion as an approach to obtain species-specific markers by PCR-RFLP analysis.