title: 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.
abstract: 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 (P35S) and MON810 construct-specific gene (MON810) 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 P35S copy number quantification. These methods should prove to be a useful tool to roughly quantify GM maize content.
authors.affiliation: Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima.
authors.forename: Akie
authors.initials: A
authors.lastname: Toyota
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Akiyama
authors.forename: Mitsunori
authors.initials: M
authors.lastname: Sugimura
authors.forename: Takahiro
authors.initials: T
authors.lastname: Watanabe
authors.forename: Kozue
authors.initials: K
authors.lastname: Sakata
authors.forename: Yuko
authors.initials: Y
authors.lastname: Shiramasa
authors.forename: Kazumi
authors.initials: K
authors.lastname: Kitta
authors.forename: Akihiro
authors.initials: A
authors.lastname: Hino
authors.forename: Muneharu
authors.initials: M
authors.lastname: Esaka
authors.forename: Tamio
authors.initials: T
authors.lastname: Maitani
mesh_terms: D004262:DNA Restriction Enzymes / Q000378:metabolism*; D018744:DNA, Plant / Q000235:genetics*; D030821:Plants, Genetically Modified; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D003313:Zea mays / Q000235:genetics*
chemical_list: D018744:DNA, Plant; D004262:DNA Restriction Enzymes