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.
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.
Toyota
Akie
A
Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima.
Akiyama
Hiroshi
H
Sugimura
Mitsunori
M
Watanabe
Takahiro
T
Sakata
Kozue
K
Shiramasa
Yuko
Y
Kitta
Kazumi
K
Hino
Akihiro
A
Esaka
Muneharu
M
Maitani
Tamio
T
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*
D018744:DNA, Plant; D004262:DNA Restriction Enzymes