Rapid Quantification Methods for Genetically Modified Maize Contents Using Genomic DNAs Pretreated by Sonication and Restriction Endonuclease Digestion for a Capillary-Type Real-Time PCR System with a Plasmid Reference Standard
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For rough quantitative analysis of genetically modified maize contents, rapid methods for measurement of the copy numbers of the cauliflower mosaic virus 35S promoter region (<I>P35S</I>) and MON810 construct-specific gene (<I>MON810</I>) using a combination of a capillary-type real-time PCR system with a plasmid DNA were established. To reduce the characteristic differences between the plasmid DNA and genomic DNA, we showed that pretreatment of the extracted genomic DNA by a combination of sonication and restriction endonuclease digestion before measurement is effective. The accuracy and reproducibility of this method for MON810 content (%) at a level of 5.0% MON810 mixed samples were within a range from 4.26 to 5.11% in the <I>P35S</I> copy number quantification. These methods should prove to be a useful tool to roughly quantify GM maize content.
- Agricultural and Biological Chemistry
Agricultural and Biological Chemistry 70(12), 2965-2973, 2006-12-23
Japan Society for Bioscience, Biotechnology, and Agrochemistry