Comparison of DNA extraction methods for entomological origin identification of honey using simple additive weighting method

Four different DNA protocols were compared to obtain the best method for DNA extraction of bees in honey samples. The efficiency was evaluated in terms of DNA concentration and purity, PCR amplification capacity targeting mitochondrial 16S rRNA gene, and execution time. The most efficient DNA extrac...

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主要な著者: Kek, Siok Peng, Chin, Nyuk Ling, Tan, Sheau Wei, Yusof, Yus Aniza, Chua, Lee Suan
フォーマット: 論文
出版事項: Blackwell Publishing Ltd 2018
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オンライン・アクセス:http://eprints.utm.my/id/eprint/86688/
http://dx.doi.org/10.1111/ijfs.13840
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要約:Four different DNA protocols were compared to obtain the best method for DNA extraction of bees in honey samples. The efficiency was evaluated in terms of DNA concentration and purity, PCR amplification capacity targeting mitochondrial 16S rRNA gene, and execution time. The most efficient DNA extraction method was selected using simple additive weighting (SAW) analysis. The DNeasy method with silica membrane spin-column format scored highest in the final ranking of SAW compared to CTAB-based, Wizard and NucleoSpin methods. It yielded extracted DNA with sufficient concentration (second highest), acceptable purity (1.5–2.0), amplifiable for 16S rRNA gene region, and required least extraction time. This recommended DNA extraction protocol suggests that genomic methodologies using a 150 bp amplicon of the 16S rRNA gene of bees enables identification of the entomological origin of honey.