In the coming years, the frequency of unauthorised genetically modified organisms (GMOs) being present
in the European food and feed chain will increase significantly. Therefore, we have developed a strategy
to identify unauthorised GMOs containing a pCAMBIA family vector, frequently present in transgenic
plants. This integrated approach is performed in two successive steps on Bt rice grains. First, the potential
presence of unauthorised GMOs is assessed by the qPCR SYBRGreen technology targeting the terminator
35S pCAMBIA element. Second, its presence is confirmed via the characterisation of the junction between
the transgenic cassette and the rice genome. To this end, a DNA walking strategy is applied using a first
reverse primer followed by two semi-nested PCR rounds using primers that are each time nested to the
previous reverse primer. This approach allows to rapidly identify the transgene flanking region and can
easily be implemented by the enforcement laboratories.
In the coming years, the frequency of unauthorised genetically modified organisms (GMOs) being present
in the European food and feed chain will increase significantly. Therefore, we have developed a strategy
to identify unauthorised GMOs containing a pCAMBIA family vector, frequently present in transgenic
plants. This integrated approach is performed in two successive steps on Bt rice grains. First, the potential
presence of unauthorised GMOs is assessed by the qPCR SYBRGreen technology targeting the terminator
35S pCAMBIA element. Second, its presence is confirmed via the characterisation of the junction between
the transgenic cassette and the rice genome. To this end, a DNA walking strategy is applied using a first
reverse primer followed by two semi-nested PCR rounds using primers that are each time nested to the
previous reverse primer. This approach allows to rapidly identify the transgene flanking region and can
easily be implemented by the enforcement laboratories.
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