Quantitative-competitive polymerase chain reaction coupled with slab gel and capillary electrophoresis for the detection of roundup ready soybean and maize.
The aim of the present study was to develop a quantitative-competitive PCR (QC-PCR) method to detect DNA from transgenic herbicide-resistant (roundup ready, RR) soybean and maize. Since no QC-PCR system for the quantification of RR maize had been published at the time of writing, a specific competitor DNA for transgenic event was developed. For the QC-PCR of RR-soybean, a commercially available competitor was employed. These internal standards were calibrated by coamplifying with mixtures containing RR-soybean and maize DNAs. The calibrated QC-PCR systems were applied to certified RR-soybean and maize flour mixtures in order to demonstrate their suitability not only for the quantification of the glyphosate resistance traits in DNA matrices, but also in practically relevant samples. In addition, a special focus of the present work was to compare the detection of QC-PCR products by slab gel and CGE with UV detection. CGE permitted the precise detection of transgenic events also below the equivalence points; while in slab gel electrophoresis, due to the low sensitivity the quantification of genetically modified DNA was allowed only at the equivalence point.
Dinelli
Giovanni
G
Department of Agro-Environmental Science and Technology, University of Bologna, Bologna, Italy. gdinelli@agrsci.unibo.it
Bonetti
Alessandra
A
Marotti
Ilaria
I
Minelli
Maurizio
M
Navarrete-Casas
Miguel
M
Segura-Carretero
Antonio
A
Fernández-Gutiérrez
Alberto
A
D002138:Calibration; D018744:DNA, Plant / Q000032:analysis*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D030821:Plants, Genetically Modified / Q000737:chemistry* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D025341:Principal Component Analysis; D013025:Glycine max / Q000737:chemistry*; D003313:Zea mays / Q000737:chemistry*
D018744:DNA, Plant