Study on genetic variability of oyster (Crassostrea iredalei, Faustino)in Peninsular Malaysia using RAPD-PCR Technique

Abstract

The random amplified polymorphic DNA (RAPD) technique was used to examine the genetic variability and relationship among individuals within and between populations of oysters (Crassostrea iredalei) from Terengganu, Kelantan, Perak and Kedah. The result of optimization of polymerase chain reaction (PCR) machine (Hybaid PCR Express, UK) showed that the best concentration of genomic DNA, magnesium chloride, dNTP- mixture, Fermentas Taq DNA Polymerase and primer for C. iredalei RAPD study were 50 ng, 3.0 mM, 0.4 mM, 2 U/|il and 0.4 |iM (10 picomoles) respectively. The machine produced clear RAPD banding patterns with an optimal annealing temperature of 36°C for 45 cycles. The genomic DNA was extracted from the oysters tissues by using phenol chloroform method. Twenty oligonucleotide primers (Kit A, Operon Technologies, Inc. Alameda, California) were screened and four primers were selected to amplify DNA from 80 samples of C. iredalei from the four populations. A total of 159 RAPD fragments (RAPDs) with 122 polymorphic fragments (76.73%) with the size ranging from 250 to 2000 base pairs (bp) were scored from the population.