In applied transgenic research as well as in agriculture there is an increasing need for high-throughput analyses of plants for genotypic selection or to identify the purity of seed stocks, e.g. for transgenic contaminations or the identification of pathogens. We developed and optimised conditions for the isolation of DNA from single seeds using an automated high-throughput protocol. Our results show that the system provided is capable of isolating DNA from any tested seed source. Furthermore, seeds remain capable of germinating during the homogenisation procedure. Quantification of endogenous and transgenic sequences by Real-Time PCR revealed that the prepared DNA is suitable in quality and quantity for multiple PCR analyses with both SYBR Green and hybridisation probe detection. The described method will be a useful tool for routine analyses like screening of large populations or the specific detection of genetically modified organisms (GMO).
There is an increasing need for high-throughput analyses of plants and food samples for the presence of specific DNA sequences, e.g. transgenic contaminations. We developed and optimized conditions for the automated isolation of DNA from several maize tissues and various edibles containing maize using the MagNA Pure LC system (Roche Applied Science). Our results show that the system provided is capable of isolating DNA from any tested source. Quantification of an endogenous gene by LightCycler real-time PCR revealed that the DNA is suitable in quality and quantity for multiple PCR analyses.
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