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Uso alternativo del colorante gelred en la tinción de ácidos nucleicos./alternative use of gelred dye in the staining nucleic acids

By Nestor Javier Orozco Clavijo, Jhoan Camilo Herrera Cañón and Jhon Fredy Betancur Pérez

Abstract

Objetivo: Determinar la reproducibilidad en la tinción del ADN en geles de agarosa mediante el colorante GelRedTM con el fin de reemplazar el uso del bromuro de etidio en la tinción de ácidos nucleicos. Materiales y Métodos: Se realizó extracción de   ADN total de sangre periférica a nueve muestras de pacientes con cáncer colorrectal, posteriormente se amplificó un fragmento de ADN de 494 pb asociado al exón 15 del gen APC y de seis fragmentos entre 150 pb y 350 pb asociados a los genes hMLH-1 y MSH-2; ambas preparaciones, ADN total y ADN amplificado fueron visualizadas mediante el uso del colorante GelRed y Bromuro de Etidio. Resultados: El uso del colorante GelRed para la tinción de ácidos nucleicos (ADN) permitió visualizarlos de forma eficiente. Conclusión: El reactivo GelRed es altamente reproducible y se recomienda usarlo incluido en las muestras de ADN, para disminuir la contaminación en los laboratorios de biología molecular. Objetive: Determine the reproducibility of DNA staining in agarose gels using the dyeGelRedTM in order to replace ethidium bromide in the use of nucleic acids staining.Materials and Methods: The extraction of the total DNA from nine samples and theamplified DNA fragment of 494 pb of exon 15 associated with the APC gene from sixfragments between 150 pb – 350 pb associated with the hMLH-1 and MSH-2 geneswas performed. Both preparations, total DNA and amplified DNA were visualized usingGelRed and Ethidium bromide dye. Results: Both systems of coloration gave verysimilar results; the DNA can be visualized correctly without any difficulty. Conclusion:The GelRed dye is highly reproducible and its use is recommended including in DNAsamples, thus reducing the contamination with dyes such as ethidium bromideDetermine the reproducibility of DNA staining in agarose gels using the dye GelRedTM in order to replace ethidium bromide in the use of nucleic acids staining. Materials and Methods: The extraction of the total DNA from nine samples and the amplified DNA fragment of 494 pb of exon 15 associated with the APC gene from sixfragments between 150 pb – 350 pb associated with the hMLH-1 and MSH-2 genes was performed. Both preparations, total DNA and amplified DNA were visualized using GelRed and Ethidium bromide dye. Results: Both systems of coloration gave very similar results; the DNA can be visualized correctly without any difficulty. Conclusion: The GelRed dye is highly reproducible and its use is recommended including in DNA samples, thus reducing the contamination with dyes such as ethidium bromide

Topics: , Electroforesis en gel de agarosa; Bromuro de Etidio; colorante,/Electrophoresis agarose gel, bromide ethidium, dyes., , Electrophoresis agarose gel; bromide ethidium; dyes.
Publisher: Universidad de Manizales
Year: 2013
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