Optimal Annealing Temperature of Yersinia enterocolitica ymoA gene primers using the Polymerase Chain Reaction Method
Muktiningsih Nurjayadi (a*), Gladys Indira Putri (a), Jefferson Lynford Declan (a), Ismaya Krisdawati (a), Dandy Akbar Juliansyah (a), Maharanianska Azzahra (a), Irvan Maulana (a), Niken K. Liman (a), Tiara Fahriza (a), Irma Ratna Kartika (a), Fera Kurniadewi (a), Dalia Sukmawati (b), Sri Rahayu (b), Vira Saamia (c), Dwi Anna Oktaviani Saputro (c), I Made Wiranatha (c), H. E. Enshay (d)(e)(f)

a) Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj^ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia
*muktiningsih[at]unj.ac.id
b) Department of Biology, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj^ari, 9th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia
c) Center Forensic Laboratory of the Criminal Investigation, Police of the Republic of Indonesia, Ciapmbuan Babakan Madang, Bogor, 1681, Indonesia
d) Institute of Bioproduct Development, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia.
e) School of Chemical and Energy Engineering, Faculty of Engineering, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia.
f) City of Scientific Research and Technology Applications, New Burg Al Arab, Alexandria, Egypt.

Abstract

The case of foodborne diseases has now become a serious global problem. Foodborne diseases occur due to contamination of pathogenic bacteria. One of the pathogenic bacteria that causes foodborne diseases is Yersinia enterocolitica. The aim of this research was 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/ micro Liter with a purity of A260/280 was 1.88. Optimization of the annealing temperature of Yersinia enterocolitica ymoA gene primers using the Gradient PCR in range of temperature between 57-61 degree Celsius 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 degree Celsius was chosen as the optimum primer annealing temperature for the ymoA gene in Yersinia enterocolitica, because only a brightness band produced on electrophoresis. This result can be continued with primers testing of the ymoA gene on meat samples using the Real-Time PCR method.

Keywords: Foodborne diseases, Polymerase Chain Reaction, Yersinia enterocolitica, ymoA gene.

Topic: Chemistry