ダイズおよびトウモロコシ抽出DNAの精製度の検討

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  • 笠間 菊子
    一般財団法人食品薬品安全センター秦野研究所
  • 小熊 恭代
    一般財団法人食品薬品安全センター秦野研究所
  • 穐山 浩
    国立医薬品食品衛生研究所
  • 鈴木 達也
    一般財団法人食品薬品安全センター秦野研究所
  • 渡辺 卓穂
    一般財団法人食品薬品安全センター秦野研究所
  • 小島 幸一
    一般財団法人食品薬品安全センター秦野研究所

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タイトル別名
  • Qualitative study of DNA extracted from soybean and maize
  • ダイズ オヨビ トウモロコシ チュウシュツ DNA ノ セイセイド ノ ケントウ

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The quality and yield of DNA extracted from soybean and maize samples were compared using two commercial DNA extraction kits, the DNeasy Plant Mini kit (Mini kit) and the GM quicker kit. Subsequent quantification of the extracted DNA samples by UV spectrophotometry and fluorometry revealed that the yields of soybean DNA extracted using the Mini kit were approximately three times higher when determined by UV spectrophotometry than when they were determined by fluorometry. Conversely, the yields of soybean DNA extracted using the GM quicker kit were only slightly higher when determined by UV spectrophotometry than when they were determined by fluorometry. However, the relative DNA yields of maize DNA samples estimated by UV spectrophotometry and fluorometry were 1.77 with the Mini kit and 1.52 with the GM quicker kit. To validate the soybean DNA yields obtained using both extraction kits, DNA samples were analyzed by agarose gel electrophoresis and real-time PCR of a reference gene. These analyses indicated that the Mini kit yields estimated by UV spectrometry were overestimated, due to more low-intensity bands and fewer gene copies being observed, compared to DNA extracted with the GM quicker kit. Conversely, the maize DNA yields obtained using the Mini and GM quicker kits showed only slight differences between the real-time PCR and agarose gel electrophoresis analyses. The extracted DNA samples were then analyzed by size-exclusion chromatography. The results showed that the soybean DNA extracted using the Mini kit contained more low-molecular weight impurities than the DNA extracted using the GM quicker kit and maize extracts obtained with both extraction kits. Therefore, it appears that the presence of low molecular weight impurities in soybean DNA extracted with the Mini kit interferes with the UV quantification of DNA.

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