Primer Design and Optimization of PCR Methods for Detecting Mixed Rat Meat in Food Samples

DOI

10.2991/978-94-6463-166-1_37How to use a DOI?

Keywords

Primer Design; PCR Optimization; Rodent ND5 Gene; Halal Detection

Abstract

Food safety is very important to support the halal tourism program. One of the problems faced by Muslim consumers is the possibility of mixing non-halal meat such as rats in meat-based food products. Molecularly we can detect the presence of rat meat in meat-based products using the PCR technique. It is necessary to design a PCR primer based on the type of rodentia that is often found in landfills, so that the detection results are more accurate. This study aims to design rodents-specific primers and optimize PCR method for detecting mixed rat meat in food samples. Primers were designed using Geneious Prime and Primer BLAST on NCBI (National Center for Biotechnology Information). The specific primers were designed on the mitochondrial region called ND5 gene (NADH dehydrogenase subunit 5), which are forward primer 5^′-GCAGTTCTCTTCATGATAYATAC-3^′ and reverse primer 5^′-GTTTCAGGCGTTGGTGTT-3^′ with a PCR product size of 919 bp. The optimization steps carried out in the PCR process include optimization of annealing temperature and primer concentration. The optimization results show that the optimum annealing temperature was 48 °C and the optimum primer concentration was 0.4 μM because it produces a thick and clear specific band.

Copyright

© 2023 The Author(s)

Open Access

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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TY  - CONF
AU  - Masnaini Masnaini
AU  - Afifatul Achyar
AU  - Moralita Chatri
AU  - Dwi Hilda Putri
AU  - Yuni Ahda
AU  - Irdawati
PY  - 2023
DA  - 2023/05/22
TI  - Primer Design and Optimization of PCR Methods for Detecting Mixed Rat Meat in Food Samples
BT  - Proceedings of the 3rd International Conference on Biology, Science and Education (IcoBioSE 2021)
PB  - Atlantis Press
SP  - 282
EP  - 289
SN  - 2468-5747
UR  - https://doi.org/10.2991/978-94-6463-166-1_37
DO  - 10.2991/978-94-6463-166-1_37
ID  - Masnaini2023
ER  -