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  • Citation:

    Kuribara, H., Shindo, Y., Matsuoka, T., Takubo, K., Futo, S., Aoki, N., et al. (2002). Novel reference molecules for quantitation of genetically modiWed maize and soybean. Journal of the AOAC International, 85, 1077–1089.

Publication type:

 peer-reviewed article

Comment:

 -

Content type:
detection

Keyword:
soybeans / maize / polymerase chain reaction / dna / plasmids / Diagenode

Publication Year:

 2002

Authority:

 AOAC

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  • Abstract:

    New quantitation methods based on a real-time polymerase chain reaction (PCR) technique were developed for 5 lines of genetically modified (GM) maize, including MON810, Event176, Bt11, T25 and GA21, and a GM soya, Roundup Ready. Oligonucleotide DNA, including specific primers and fluorescent dye-labelled probes, were designed for PCRs. Two plasmids were constructed as reference molecules (RMs) for the detection of GM maize and GM soya. The molecules contain the DNA sequences of a specific region found in each GM line, universal sequences used in various GM lines, such as cauliflower mosaic virus 35S promoter and nopaline synthase terminator, and the endogenous DNA sequences of maize or soya. By using these plasmids, no GM maize and GM soya were required as reference materials for the qualitative and quantitative PCR technique. Test samples containing 0, 0.10, 0.50, 1.0, 5.0 and 10% GM maize or GM soya were quantitated. At the 5.0% level, the bias (mean-true value) ranged from 2.8 to 19.4% and the relative standard deviation was < 5.2%. These results show that our method involving the use of these plasmids as RMs is reliable and practical for quantitation of GM maize and GM soya.