Open AccessNovel primer targeting the mitochondrial NADH dehydrogenase subunit 4 (ND4) and NADH dehydrogenase subunit 5 (ND5) for detection of porcine (Sus scrofa) DNA fragments in food products for halal authentication
Author(s) -
Joni Kusnadi,
P Mahardita,
K U Al-Awwaly,
Estri Laras Arumingtyas
Publication year - 2021
Publication title -
iop conference series. earth and environmental science
Language(s) - English
Resource type - Journals
eISSN - 1755-1307
pISSN - 1755-1315
DOI - 10.1088/1755-1315/924/1/012004
Subject(s) - nadh dehydrogenase , primer (cosmetics) , microbiology and biotechnology , protein subunit , biology , mitochondrial dna , alcohol dehydrogenase , dna , gene , polymerase chain reaction , biochemistry , chemistry , enzyme , organic chemistry
In this research, a pair primer targeting NADH dehydrogenase subunit 4 (ND4 ) gene of porcine ( Sus scrofa ) mt-DNA was designed and tested for its specificity. The ND4 primer was compared for its robustness against an established primer pair designed to amplify the NADH dehydrogenase subunit 5 (ND5 ) gene. The samples include non-halal meat animals (dog, porcine, and rat) and halal meat animals (chicken, cow, and horse). DNA from raw meat was isolated by modified Chloroform Isoamyl-Alcohol method and then was analyzed quantitatively with NanoDrop™ spectrophotometer followed by amplification using PCR technique. The amplification results proved that the two pairs of primers were specific, resulting in amplification of 120 bp DNA target for ND4 and 227 bp for ND5 . It can be concluded that the two primers can differentiate between halal and non-halal animals. However, to determine the sensitivity of each primer, further research is needed.