From Germs to Mammals in Aqua
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Genetics of Aquatic Organisms 2017, Vol 1, Num, 1     (Pages: 21-26)

Impact of Enzymes and Primers on the PCR Amplification of Some Goatfishes

Taha Soliman 1-2 ,Eman M. Abbas 2 ,Reda M. Fahim 2

1 Okinawa Institute of Science and Technology, 1919-1 Tancha, Onna-son, Kunigami, Okinawa 904-0495, Japan
2 National Institute of Oceanography and Fisheries, Kayet-Bay, El-Anfoushy, 21556 Alexandria, Egypt
DOI : 10.4194/2459-1831-v1_1_04 Viewed : 8448 - Downloaded : 5021 The polymerase chain reaction (PCR) is one of the most powerful techniques in molecular biology. In DNA isolation, substances co-extracted from biological samples, such as lipids, polysaccharides, and humic acids, influence amplification of the target gene and act as PCR inhibitors. Here, two instruments were used to measure concentrations and purities of DNA extracted from three species of the family Mullidae (Mullus barbatus, Linnaeus, 1758; Mullus surmuletus, Linnaeus, 1758; Upeneus moluccensis, Bleeker, 1855). In order to investigate PCR amplification differences between these species of goatfishes and to optimize protocols for PCR amplification, two PCR enzymes with different annealing temperatures were tested by amplifying the mitochondrial COI gene. The high lipid level of M. surmuletus species acted as a polymerase chain reaction inhibitor. The findings presented herein will enable other researchers to choose enzymes and primers appropriate for their studies instead of merely adjusting PCR annealing temperatures. Keywords : Mediterranean Sea, Egypt, family Mullidae, PCR