Selection of the most suitable method for the extraction of DNA from foods and feeds for species identification
2010
Nesvadbova, M.,Mendelova Zemedelska a Lesnicka Univ., Brno (Czech Republic). Ustav Morfologie, Fyziologie a Genetiky Zvirat | Knoll, A.,Mendelova Zemedelska a Lesnicka Univ., Brno (Czech Republic). Ustav Morfologie, Fyziologie a Genetiky Zvirat | Vasatkova, A.,Mendelova Zemedelska a Lesnicka Univ., Brno (Czech Republic). Ustav Morfologie, Fyziologie a Genetiky Zvirat
High quality and purity of DNA isolated from food and feed is essential for species identification. The efficiency of eight different methods for DNA isolation was investigated. Raw chicken meat, ham, sausages, tinned lunch meat, pate, tinned feeds for dogs, complete granulated feeds for dogs and chicken flour were analysed. The following kits were employed: NucleoSpin Food (Marchery-Nagel), Wizard Genomic DNA Purification Kit (Promega), Invisorb Spin Food Kit I (Invitek), Wizard SV Genomic DNA Purification System (Promega), JetQuick Tissue DNA Spin Kit (Genomed), RNA Blue (Top-Bio), JetQuick Blood and Cell Culture Kit (Genomed), QIAamp DNA Mini Kit and QIAamp DNA Blood Mini Kit (Qiagen). Gel agarose electrophoresis for primary verification of DNA quality was performed. Quantities and qualities of the isolates were subsequently assessed by the spectrophotometer Nanodrop 2000 (Thermo Scientific). DNA quality was also verified by PCR. Differences between the isolated DNA from tinned products and meat, ham, sausage, granulated dog feed and chicken flour were found. In tinned food and feed, the DNA was more degraded, DNA content and DNA purity was lower and also PCR amplification was the most difficult. Overall DNA yield and quality have important influence on PCR products amplification. The best results were obtained with NucleoSpin Food and JetQuick Tissue DNA Spin Kit. Our results depended on different food or feed and different isolation system.
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