English   español  
Por favor, use este identificador para citar o enlazar a este item: http://hdl.handle.net/10261/50320
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

Simultaneous detection of genetically modified organisms by multiplex ligation-dependent genome amplification and capillary gel electrophoresis with laser-induced fluorescence

AutorGarcía-Cañas, Virginia ; Mondello, Monica; Cifuentes, Alejandro
Palabras claveCGE-LIF
DNA amplification
Food analysis
Genetically modified organisms
Transgenic maize
Fecha de publicaciónjul-2010
EditorWiley-VCH
CitaciónElectrophoresis 31(13): 2249-2259 (2010)
ResumenIn this work, an innovative method useful to simultaneously analyze multiple genetically modified organisms is described. The developed method consists in the combination of multiplex ligation-dependent genome dependent amplification (MLGA) with CGE and LIF detection using bare-fused silica capillaries. The MLGA process is based on oligonucleotide constructs, formed by a universal sequence (vector) and long specific oligonucleotides (selectors) that facilitate the circularization of specific DNA target regions. Subsequently, the circularized target sequences are simultaneously amplified with the same couple of primers and analyzed by CGE-LIF using a bare-fused silica capillary and a run electrolyte containing 2-hydroxyethyl cellulose acting as both sieving matrix and dynamic capillary coating. CGE-LIF is shown to be very useful and informative for optimizing MLGA parameters such as annealing temperature, number of ligation cycles, and selector probes concentration. We demonstrate the specificity of the method in detecting the presence of transgenic DNA in certified reference and raw commercial samples. The method developed is sensitive and allows the simultaneous detection in a single run of percentages of transgenic maize as low as 1% of GA21, 1% of MON863, and 1% of MON810 in maize samples with signal-to-noise ratios for the corresponding DNA peaks of 15, 12, and 26, respectively. These results demonstrate, to our knowledge for the first time, the great possibilities of MLGA techniques for genetically modified organisms analysis.
Descripción11 páginas, 5 figuras, 2 tablas.-- El pdf del artículo es la versión revisada de autor.
Versión del editorhttp://dx.doi.org/10.1002/elps.201000078
URIhttp://hdl.handle.net/10261/50320
DOI10.1002/elps.201000078
ISSN0173-0835
E-ISSN1522-2683
Aparece en las colecciones: (IFI) Artículos
Ficheros en este ítem:
Fichero Descripción Tamaño Formato  
Simultaneous detection.pdf716,66 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.