Polymerase Chain Reaction-Restriction Fragment Length Polymorphism of Mitochondrial 12S rRNA gene. A simple method for identification of Poultry Meat Species.

Abstract

Chicken (Gallus gallus), duck (Anas platyrhynchos), turkey (Meleagris gallopavo), guinea fowl (Numida me- leagris)andquail(Coturnixjaponica)arethecommonpoultryspeciesconsumedasmeatthroughouttheworld. Inthiswork,amoleculartechniquehasbeendevelopedforidentificationanddifferentiationofmeatoriginating from these species. This tool helps in detection of misrepresentation of different poultry meats. The technique involves the extraction of DNA from the given sample, polymerase chain reaction (PCR) amplification of mitochondrial 12S rRNA gene using universal primers, restriction analysis with selected restriction enzymes, followed by identification of meat species based on restriction fragment length polymorphism (RFLP) pattern. In this study, we used HinfI, Mph1103I, MvaI, and Eco47I to identify and differentiate to poultry species referred to above. This species identification technique has also been applied successfully to processed meat products including those cooked at 120◦C for 30min. Simplicity of interpretation of results combined with versatility makes this a convenient and appropriate technique in the hands of meat analysts for identifying poultry meat species. Keywords: 12S rRNA gene, meat species, PCR, poultry, RFLP Abbreviations: bp, base pairs; dNTP, deoxynucleotide triphosphate; DNAase, deoxyribonuclease; DTT, Dithiothreitol; EDTA, ethylenediaminetetraacetate; mt, mitochondrial; PCR, polymerase chain reaction; RAPD, random amplified polymorphic DNA; RE, restriction enzyme; RFLP, restriction fragment length polymorphism; RNase, riboendonuclease; SDS, sodium dodecyl sulphate; TE buffer, Tris-EDTA buffer