English   español  
Por favor, use este identificador para citar o enlazar a este item: http://hdl.handle.net/10261/108267
COMPARTIR / IMPACTO:
Estadísticas
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL
Exportar a otros formatos:
Título

A new PCR-CGE (size and color) method for simultaneous detection of genetically modified maize events

AutorNadal, Anna ; Coll, Anna; La-Paz, José Luis ; Esteve, Teresa; Pla, Maria
Palabras claveFluorescence
Genetically modified organism
Maize
Multiplex PCR
CE
Fecha de publicaciónoct-2006
EditorJohn Wiley & Sons
CitaciónElectrophoresis 27(19): 3879-3888 (2006)
ResumenWe present a novel multiplex PCR assay for simultaneous detection of multiple transgenic events in maize. Initially, five PCR primers pairs specific to events Bt11, GA21, MON810, and NK603, and Zea mays L. (alcohol dehydrogenase) were included. The event specificity was based on amplification of transgene/plant genome flanking regions, i.e., the same targets as for validated real-time PCR assays. These short and similarly sized amplicons were selected to achieve high and similar amplification efficiency for all targets; however, its unambiguous identification was a technical challenge. We achieved a clear distinction by a novel CGE approach that combined the identification by size and color (CGE-SC). In one single step, all five targets were amplified and specifically labeled with three different fluorescent dyes. The assay was specific and displayed an LOD of 0.1% of each genetically modified organism (GMO). Therefore, it was adequate to fulfill legal thresholds established, e.g., in the European Union. Our CGE-SC based strategy in combination with an adequate labeling design has the potential to simultaneously detect higher numbers of targets. As an example, we present the detection of up to eight targets in a single run. Multiplex PCR-CGE-SC only requires a conventional sequencer device and enables automation and high throughput. In addition, it proved to be transferable to a different laboratory. The number of authorized GMO events is rapidly growing; and the acreage of genetically modified (GM) varieties cultivated and commercialized worldwide is rapidly increasing. In this context, our multiplex PCR-CGE-SC can be suitable for screening GM contents in food. © 2006 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
Versión del editorhttp://dx.doi.org/10.1002/elps.200600124
URIhttp://hdl.handle.net/10261/108267
DOI10.1002/elps.200600124
Identificadoresdoi: 10.1002/elps.200600124
issn: 0173-0835
Aparece en las colecciones: (IBMB) Artículos
Ficheros en este ítem:
Fichero Descripción Tamaño Formato  
accesoRestringido.pdf15,38 kBAdobe PDFVista previa
Visualizar/Abrir
Mostrar el registro completo
 

Artículos relacionados:


NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.