Identification of dna marker in tiger grouper (Epinephelus fuscoguttatus) from Kedah population

Abstract

A study to clone and identify DNA marker in Tiger Grouper (Epinephelus fuscoguttatus) in Kedah population was carried out. In this study, DNA from four population of Tiger grouper that from Bali, Kedah, Langkawi and Sabah was extracted from tissue muscles and then used as a template for Polymerase Chain Reaction (PCR) amplification using RAPD-PCR technique. In this PCR Amplification, three types of primer that OPA-01, OPA-11 and OPA-20 had used to produce DNA profile from these four population. Result showed that only one polymorphic fragment sized 504 hp exist from sample E. fuscoguttatus from Kedah population by using primer OPA-01. After that, this fragment was isolated and purified for cloning purpose. In cloning, this fragment was ligated using cloning vector pTZ57R/7 sized 2886 bp and transformation had done using bacteria cell E. coli. Cloning recombinant had successfully got by used plasmid sized -3300 bp. From the result of BLAST showed that there have no significant similarity found and that's mean there have no reported that this sequence have found yet and also this result showed and also maybe it specific for species E. fuscoguttatus. It expected because sequence of E. fuscoguttatus just 0.00002% only compared with model organism zebra fish (Danio rerio ). The future study about this experiment that can design primer according to the cloned sequence and this could be easily and effectively to determine between populations using PCR technique.