PCR-RFLP analysis has been applied to the identification of meats from red deer (Cervus elaphus),
fallow deer (Dama dama), roe deer (Capreolus capreolus), cattle (Bos taurus), sheep (Ovis aries),
and goat (Capra hircus). PCR amplification was carried out using a set of primers flanking a conserved
region of ∼712 base pairs from the mitochondrial 12S rRNA gene. Restriction site analysis based on
sequence data from this DNA fragment permitted the selection of MseI, MboII, BslI, and ApoI
endonucleases for species identification. The restriction profiles obtained when amplicons were
digested with the chosen enzymes allowed the unequivocal identification of all domestic and game
meat species analyzed in the present work.
Keywords: Game meat; species identification; 12S rRNA gene; PCR-RFLP
A polymerase chain reaction (PCR) assay was developed for the specific identification of cows' milk in sheep's and goats' milk by using primers targeting the mitochondrial 12S rRNA gene. The use of a forward primer complementary to a conserved DNA sequence, along with a reverse primer specific for cow, yielded a 223-bp fragment from cows' milk DNA, whereas no amplification signal was obtained in sheep's and goats' milk DNA. The technique was applied to raw, pasteurized, and sterilized milk binary mixtures of cow-sheep and cow-goat, enabling the specific detection of cows' milk with a good sensitivity threshold (0.1%). The proposed PCR assay represents a rapid and straightforward method applicable to the authentication of milk and other dairy products in routine analysis.
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