Optimal annealing temperature of Yersinia enterocolitica ymoA gene primers using the polymerase chain reaction method

The case of foodborne diseases has now become a serious global problem. Foodborne diseases occur due to the contamination of pathogenic bacteria. One of the pathogenic bacteria that cause foodborne diseases is Yersinia enterocolitica. This research aimed to determine the annealing temperature of Yersinia enterocolitica ymoA gene primers using the PCR method to ensure that the primers can amplify ymoA gene fragments in Yersinia enterocolitica. In this research, the concentration of DNA as a template used was 70.5 ng/µL with a purity of A260/280 1.88. Optimization of the annealing temperature of Yersinia enterocolitica ymoA gene primers using the Gradient PCR in a range of temperatures between 53°C - 62°C based on the melting temperature value of the ymoA primers. The results of the primers design of the ymoA gene were confirmed to amplify the DNA fragment of Yersinia enterocolitica with an amplicon length of 185 bp. Based on the results, the temperature of 60°C was chosen as the optimum primer annealing temperature for the ymoA gene in Yersinia enterocolitica because only a brightness band was produced from the electrophoresis. This result can be continued with primers testing the ymoA gene on meat samples using the Real-Time PCR method.