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Real-time loop-mediated isothermal amplification for the CaMV-35S promoter as a screening method for genetically modified organisms

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Abstract

A method has been developed to detect genetically modified organisms (GMOs). The detection method is based on the loop-mediated isothermal amplification (LAMP) reaction. Cauliflower mosaic virus 35S (CaMV-35S) promoter gene, a widespread genetic element, was amplified by a set of LAMP primers. One of the characteristics of the LAMP method is its ability to synthesize an extremely large amount of DNA. Accordingly, a large amount of byproduct, pyrophosphate ion, is produced, yielding a white precipitate of magnesium pyrophosphate in the LAMP reaction mixture. The measurement of the turbidity of the reaction mixture allows easy detection of amplification of CaMV-35S promoter gene without the need for gel electrophoresis. With this new technique we analyzed Roundup Ready soybean, and found GMO content ranging from 0.5% to 5%.

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Authors and Affiliations

  1. Aichi-Pref. Agricultural Research Center, 1–1 Sagamine, Yazako, Nagakute, Aichi, 480–1193, Japan

    Shiro Fukuta, Yuko Mizukami, Akira Ishida, Junichi Ueda, Mayumi Hasegawa, Izumi Hayashi, Minako Hashimoto & Michio Kanbe

Authors
  1. Shiro Fukuta
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  2. Yuko Mizukami
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  3. Akira Ishida
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  4. Junichi Ueda
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  5. Mayumi Hasegawa
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  6. Izumi Hayashi
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  7. Minako Hashimoto
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  8. Michio Kanbe
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Correspondence to Shiro Fukuta.

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Fukuta, S., Mizukami, Y., Ishida, A. et al. Real-time loop-mediated isothermal amplification for the CaMV-35S promoter as a screening method for genetically modified organisms. Eur Food Res Technol 218, 496–500 (2004). https://doi.org/10.1007/s00217-003-0862-5

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  • DOI: https://doi.org/10.1007/s00217-003-0862-5

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