Untitled

Abstract

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.