DEVELOPMENT OF TESTING METHOD FOR PORK CONTAMINATION IN CORNED BEEF USING POLYMERASE CHAIN REACTION (PCR) SPECIFIC PRIMER TECHNIQUE; PENGEMBANGAN METODE UJI KONTAMINASI DAGING BABI PADA KORNET SAPI MENGGUNAKAN TEKNIK POLYMERASE CHAIN REACTION (PCR) PRIMER SPESIFIK
Rachmadian, Fayme, Tri Joko Raharjo
2015 | Skripsi | FMIPAAdulteration of meat and processed meat product with substitution of pork is a problem in the industry. A test for assessing pork adulteration in corned beef using polymerase chain reaction (PCR) specific primer detection of a conserved region in the mitochondrial (mt) ND5 gene has been performed. The research was aimed to develop identification method of pork contamination in corned beef based on DNA test that can be applied for Halal authentication. This study consisted of two stages, namely method performance test (specificity test and determination of cut off limit) and application of method on commercial corned beef. DNA of corned samples were extracted using NucleoSpin Food kit then analyzed by UV spectrophotometry and electrophoresis, followed by PCR amplification of ND5 genes fragment using specific primer of ND5 forward primer (5’-CAT TCG CCT CAC TCA CAT TAAC-3’) and the ND5 reverse primer (5’-AAG AGA GAG TTC TAC GGT CTG TAG-3’). The same procedures were performed on method performance test and the application of method. Extracted DNA from the corned samples were found to be good quality and produced clear PCR products on the amplification of ND5 gene of 227 base pairs (bp) in size specific for pork species. The performance test showed that the assay has high performance and could be used to detected pork contamination in corned beef with cut off limit to the level of 1% (w/w). It was a potentially reliable and suitable technique in routine food analysis for halal authentication. The analysis of four commercial corned beef showed no pork contamination in all samples.
Kata Kunci : PCR; ND5; Corned; Pork; Halal Analysis
