Specific Primer Design of COI Gene and Its Potential Application for Species Identification of Meats

Abstract

Cytochrome oxidase I gene, COI, is known as a mitochondrial gene encoding cytochrome oxidase I protein. The aim of this study was to design specific primers for bovine, chicken, and porcine and its application for species identification containing in raw meats using PCR. A set of specific primers has been designed using primer3. A universal forward primer and three specific reverse primers have been synthesized and the PCR technique has been carried out to test their specificity using a DNA template extracted from three meat species, i.e. bovine, chicken, and porcine. The PCR condition was optimized with similar annealing temperatures at 64°C. The PCR product was then visualized using a 2% agarose gel under the UV light and sequenced to know its nucleotides. The results showed that primer pairs were perfectly working to amplify specific target sites for bovine, chicken, and porcine. They were indicated by 263 bp for bovine, 596 bp for chicken, and 168 bp for porcine. Besides, sequence analyses showed that they were a hundred percent similar to reference species. This result suggested that the primer set of COI genes designed in this study may be a powerful tool to identify species in meats using multiplex-PCR.