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All Journal Jurnal Pembelajaran dan Biologi Nukleus
Siboro, Dewi Pratiwi Purba
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Education Environmental Science Immunology & microbiology Languange, Linguistic, Communication & Media

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Development of Species-Specific Primers Targeting Mitochondrial Cyt b Gene for Porcine DNA Detection in Halal Authentication via Polymerase Chain Reaction (PCR) Malau, Jekmal; Kasasiah, Ahsanal; Zahra, Aliya Azkia; Irwansyah, Silvana Lestari; Siboro, Dewi Pratiwi Purba
JURNAL PEMBELAJARAN DAN BIOLOGI NUKLEUS Vol 11, No 2: Jurnal Pembelajaran Dan Biologi Nukleus June 2025
Publisher : Universitas Labuhanbatu

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.36987/jpbn.v11i2.6799

Abstract

Background: The detection of porcine DNA is critical to ensuring adherence to halal standards, particularly in food and pharmaceutical products. This study aimed to design and validate species-specific primers targeting the mitochondrial Cyt b gene of Sus scrofa for porcine DNA identification. Using in silico tools such as NCBI, Primer3Plus, SnapGene, Mega and NetPrimer, four primer pairs were designed and assessed for specificity and efficiency. Methodology: Laboratory validation involved PCR amplification and bi-directional Sanger sequencing. Findings: The findings demonstrated that primers 2F/2R and 3F/3R successfully amplified the target DNA, producing amplicons of 514 bp and 456 bp, respectively. The primers exhibited high specificity, with no amplification observed in non-target DNA samples, including chicken and beef. Sanger sequencing confirmed that the amplified products corresponded to the Cyt b gene sequence of Sus scrofa with 100 % similarity, as validated through BLAST analysis. This study presents an accurate and dependable molecular method for detecting porcine DNA, with valuable applications in halal authentication and molecular diagnostics. Contribution: The developed primers offer an effective tool for accurately identifying porcine-derived components, addressing the critical demand for species-specific DNA detection to support halal compliance.
Co-Authors Ahsanal Kasasiah Irwansyah, Silvana Lestari Jekmal Malau Zahra, Aliya Azkia