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Detection of land animal remains in fish meals by the polymerase chain reaction-restriction fragment length polymorphism technique
Authors:Santaclara Francisco J  Espiñeira Montserrat  Cabado Ana G  Vieites Juan M
Institution:Area of Molecular Biology and Biotechnology, ANFACO-CECOPESCA, Vigo, 36310 Pontevedra, Spain. fransanta@anfaco.es
Abstract:In the present study a technique was developed with the aim of guaranteeing the composition and security of fish meals, since it allows verification of whether these meals contain land animal remains. The method is based on polymerase chain reaction (PCR) and length polymorphism, followed by a restriction fragment length polymorphism (RFLP). Specific primers for every species were designed and calibrated, generating exclusively a PCR product with a specific size when DNA for each species was present in the sample. This technique allows the detection of land animal remains in fish meals, specifically cow, chicken, pig, horse, sheep, and goat. The identity of the PCR products can be confirmed by RFLP analysis using only one restriction enzyme. The selected restrictase generated one characteristic restriction profile for every species included in this study. The detection limit of this method was calculated by using mixtures of fish meals in different proportions and meal that exclusively contained remains of one of these land species studied. The analytical strategy herein proposed was applied to fish and meat meals, giving good results, both in the analyzed standards and in commercial samples.
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