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 | Zendy

Akie Toyota | Zendy; Hiroshi Akiyama | Zendy; Mitsunori SUGIMURA | Zendy; Takahiro Watanabe | Zendy; Kozue Sakata | Zendy; Yuko Shiramasa | Zendy; Kazumi Kitta | Zendy; Akihiro Hino | Zendy; Muneharu Esaka | Zendy; Tamio Maitani | Zendy

Open Access

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

Author(s) -

Akie Toyota,

Hiroshi Akiyama,

Mitsunori SUGIMURA,

Takahiro Watanabe,

Kozue Sakata,

Yuko Shiramasa,

Kazumi Kitta,

Akihiro Hino,

Muneharu Esaka,

Tamio Maitani

Publication year - 2006

Publication title -

bioscience biotechnology and biochemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.509

H-Index - 116

eISSN - 1347-6947

pISSN - 0916-8451

DOI - 10.1271/bbb.60366

Subject(s) - restriction enzyme , genomic dna , biology , genetically modified maize , plasmid , dna , endonuclease , microbiology and biotechnology , cauliflower mosaic virus , dna extraction , polymerase chain reaction , chromatography , gene , chemistry , genetics , genetically modified crops , transgene

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.

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