Identification of Meat Species by PCR-RFLP Method using Single Set of Degenerative Primers

Abstract

Due to religious and social concerns, identification of meat and meat products is a matter of great interest for general public. DNA-based identification methods are ideal and reliable. DNA was extracted from muscle tissues of cow, buffalo, goat, donkey and a dog. A set of degenerative primers was designed for Polymerase Chain Reaction (PCR) of the cytochrome b gene fragment. All PCR was accomplished at the same reaction conditions. Amplified products had a length of 400 bp, which was our estimated length. After the sequencing of these amplicons, In silico restriction analysis was performed for each specie. Restriction endonuclease TfiI was selected for further analysis. Restriction fragment length polymorphism (RFLP) was performed at the same reaction conditions for all species. RFLP analysis produced species specific restriction profiles. Comparing these profiles, we have identified the species origin of meat unambiguously.