PCR Optimization and PCR-RFLP Analysis of Ambal Biasa (Solen regularis) from Kuching Bay Areas

• Main Author: Crocker, Melinda Alexandria
• Format: Final Year Project Report
• Language: English
• Published: Universiti Malaysia Sarawak (UNIMAS) 2007
• Subjects: SH Aquaculture. Fisheries. Angling
• Online Access: http://ir.unimas.my/id/eprint/27683/3/Melinda.pdf; http://ir.unimas.my/id/eprint/27683/

This study was conducted to obtain a baseline data for molecular studies of 'ambal biasa' (Solen regularis) in Kuching Bay area. Samples were collected from tlu·ee sites namely Bako, Asajaya and Buntal in the month of September 2006. There were 38 individuals of S. regularis that were used in this study. The modified CT AB protocol was used in utilizing the total genomic DNA ofthe S. regularis. PCR had been carried out to amplify the l6S mtDNA gene and ITS··1 gene region. Primers 16Sar-L (Forward) and 16Sbr-H (Reverse) were used for l6S mtDNA while ETTS-2 (Forward) and ITS-l r (Reverse) were used for ITS-l gene. The PCR annealing temperature acquired for gene 16S mtDNA was at 45°Celsius whereas the optimal temperature was at 54°Celsius for ITS-l gene. Four enzymes had been used in PCRRFLP experiment namely Alu1, BamH1, Mps1 , Sau3A and BSlz1. However, only RE Alul showed positive result.