Development of a Multiplex Ligation Dependent Probe Amplification MLPA Module for Simultaneous Detection of Five Genetically Modified Rapeseed EventsReportar como inadecuado




Development of a Multiplex Ligation Dependent Probe Amplification MLPA Module for Simultaneous Detection of Five Genetically Modified Rapeseed Events - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

For the official surveillance of genetically modified crops, efficientand simple detection methods need to be on hand to implement the strictrequirements regarding approval, labelling and traceability determined by theEuropean Union. Therefore, a multiplex ligation dependent probe amplificationMLPA module was developed for simultaneous detection of five geneticallymodified rapeseed events MS8, RF3, GT73, Falcon GS40-90 and T45. Probes weredesigned and concentrations were adapted in order to obtain high sensitivityand specificity. This MLPA module was validated using certified referencematerials and its applicability was tested analyzing routine honey, mustard andrapeseed samples. The limit of detection was determined by analyzing a dilutionseries n = 16 for each concentration of respective transgenic DNA. Afteroptimization, the MLPA revealed limits of detection between 10 to 50 copies ofthe transgene DNA-assay. The method proved to be sensitive and highlyreproducible. When analyzing routine samples, results obtained applying theMLPA module were similar compared to real-time PCR.

KEYWORDS

MLPA, Genetically Modified Organism, Rapeseed, Capillary Electrophoresis, Detection

Cite this paper

Guertler, P. , Goerlich, O. and Busch, U. 2014 Development of a Multiplex Ligation Dependent Probe Amplification MLPA Module for Simultaneous Detection of Five Genetically Modified Rapeseed Events. Agricultural Sciences, 5, 530-539. doi: 10.4236-as.2014.56055.





Autor: Patrick Guertler, Ottmar Goerlich, Ulrich Busch

Fuente: http://www.scirp.org/



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