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http://dx.doi.org/10.5851/kosfa.2015.35.3.382

A Simultaneous Analytical Method for Duplex Identification of Porcine and Horse in the Meat Products by EvaGreen based Real-time PCR  

Sakalar, Ergun (Department of Bioengineering, Faculty of Engineering, Canakkale Onsekiz Mart University)
Ergun, Seyma Ozcirak (Department of Bioengineering, Faculty of Engineering, Canakkale Onsekiz Mart University)
Akar, Emine (Department of Bioengineering, Faculty of Engineering, Canakkale Onsekiz Mart University)
Publication Information
Food Science of Animal Resources / v.35, no.3, 2015 , pp. 382-388 More about this Journal
Abstract
A duplex real-time polymerase chain reaction (PCR) based assay for the detection of porcine and horse meat in sausages was designed by using EvaGreen fluorescent dye. Primers were selected from mitochondrial 12S rRNA and 16S rRNA genes which are powerful regions for identification of horse and porcine meat. DNA from reference samples and industrial products was successfully extracted using the GIDAGEN® Multi-Fast DNA Isolation Kit. Genomes were identified based on their specific melting peaks (Mp) which are 82.5℃ and 78℃ for horse and porcine, respectively. The assay used in this study allowed the detection of as little as 0.0001% level of horse meat and 0.001% level of porcine meat in the experimental admixtures. These findings indicate that EvaGreen based duplex realtime PCR is a potentially sensitive, reliable, rapid and accurate assay for the detection of meat species adulterated with porcine and horse meats.
Keywords
duplex real-time PCR; EvaGreen; horse; porcine; unspecific amplicon;
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