Open AccessSpecific Primer Design of COI Gene and Its Potential Application for Species Identification of Meats
Author(s) -
T. Khoirinisah,
A. Fadhila,
Tommy Wibowo,
Lilik Retna Kartikasari,
Muhammad Cahyadi
Publication year - 2020
Publication title -
iop conference series. earth and environmental science
Language(s) - English
Resource type - Journals
eISSN - 1755-1307
pISSN - 1755-1315
DOI - 10.1088/1755-1315/478/1/012040
Subject(s) - primer (cosmetics) , biology , multiplex polymerase chain reaction , mitochondrial dna , gene , microbiology and biotechnology , genetics , agarose gel electrophoresis , polymerase chain reaction , computational biology , chemistry , organic chemistry
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.