Genomic DNA of 13 fish (n=13) species consist of four freshwater which were catfish (Clarias gariepinus), shark catfish (Pangasius larnaudii), tilapia (Oreochromis mossambicus), perch (Lates calcarifer) and nine marine species which were black pomfret (Parastromateus niger), anchovy (Stolephorus commersonii), mabong (Rastrelliger kanagurta), red snapper (Lutjanus erythropterus), herring (Chirocentrus dorab), ray fish (Himantura gerrardii), sardine (Decapterus macrosoma), mackerel (Euthynnus affinis) and tuna (Thunnus tuna) were differentiated using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Seven endonucleases of AluI, BsaJI, HaeIII, HindIII, HinfI, MboI and MboII were examined for the ability to digest cyt b amplicon from each species. Genomic DNA of pork (Sus scrofa domestica) were differentiated from fishes by comparing the digestion patterns produced by similar amplified region and enzymes used. In the present study, it was demonstrated that fishes and pork DNA genome were successfully differentiated using all endonucleases except for HindIII. Thus, PCR-RFLP analysis was found useful for future pork DNA detection in fish products.
A method of PCR-restriction fragment length polymorphism (RFLP) has been utilized to differentiate the mitochondrial genes of pork and wild boar meat (Sus scrofa). The amplification PCR products of 359 bp and 531 bp were successfully amplified from the cyt b gene of these two meats. The amplification product of pork and wild boar using mt-12S rRNA gene successfully produced a single band with molecular size of 456 bp. Three restriction endonucleases (AluI, HindIII and BsaJI) were used to restrict the amplification products of the mitochondrial genes. The restriction enzymes of AluI and BsaJI were identified as potential restriction endonucleases to differentiate those meats. HindIII enzyme was unable to restrict the PCR product of both meats. The genetic differences within the cyt b gene among the two meats were successfully confirmed by PCR-RFLP analysis.