PENGEMBANGAN METODE AUTENTIKASI HALAL PADA KORNET SAPI DENGAN POLYMERASE CHAIN REACTION-RESTRICTION FRAGMENT LENGTH POLYMORPHISM (PCR-RFLP) GEN cytB DNA MITOKONDRIA; DEVELOPMENT OF HALAL AUTHENTICATION METHOD ON CORNED BEEF USING POLYMERASE CHAIN REACTION-RESTRICTION FRAGMENT LENGTH POLYMORPHISM (PCR-RFLP) cytB GENE MITOCHONDRIAL DNA

Fardani, Dwi Rizki A, Tri Joko Raharjo

2015 | Skripsi | FMIPA

Analysis of pork content in corned beef using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) cytB gene mitochondrial DNA has been performed. The aim of this study was to develop a pork contamination method in corned beef based on DNA test that can be used as halal authentication method. This study consisted of two stages, i.e. validation and application of method. Validation of method included specificity and sensitivity tests which were started with isolation of DNA using phenol-CIAA. DNA isolate was used as template in DNA amplification process by using primers cytB forward L14841 (5'-CCATCCAACATCTCAGCATGATGAAA-3') and reverse H15149 (5'-GCCCCTCAGAATGATATTTGTCCTCA-3'). Then, the amplicon DNA was restricted by enzyme BseDI at 55 °C for 4 hours. The result showed that amplification process has occured on DNA fragment with length 359 bp. Restriction of DNA amplicon showed the cutting pattern on 228 and 131 bp in pork and did not in beef. Validation of method showed that PCR-RFLP method was specific to detect pork contamination with limit of detection to the level of 1%. Four commercial corned beef samples were tested negative for pork contamination.

Kata Kunci : halal; corned beef; cytB; PCR-RFLP