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Directed Covalent Immobilization of Aminated DNA Probes on Aminated Plates

Fuentes, Manuel ; Mateo, Cesar ; García, Lucia ; Tercero, Juan C ; Guisán, José M ; Fernández-Lafuente, Roberto

Biomacromolecules, 2004-05, Vol.5 (3), p.883-888 [Periódico revisado por pares]

Washington, DC: American Chemical Society

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Directed Covalent Immobilization of Aminated DNA Probes on Aminated Plates
Autor: Fuentes, Manuel ; Mateo, Cesar ; García, Lucia ; Tercero, Juan C ; Guisán, José M ; Fernández-Lafuente, Roberto
Assuntos: Adsorption ; Amines - chemistry ; Applied sciences ; Base Sequence ; Biological and medical sciences ; Biotechnology ; DNA Primers ; DNA Probes ; Exact sciences and technology ; Fundamental and applied biological sciences. Psychology ; Immobilization of enzymes and other molecules ; Immobilization techniques ; Methods. Procedures. Technologies ; Natural polymers ; Physicochemistry of polymers ; Starch and polysaccharides
É parte de: Biomacromolecules, 2004-05, Vol.5 (3), p.883-888
Notas: ObjectType-Article-2
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Descrição: A new protocol that enables the immobilization of DNA probes on aminated micro-titer plates activated with aldehyde-dextran via an amino group artificially introduced in the 3‘ end of the oligonucleotide probe is reported in this work. The method is based on the use of hetero-functional-dextran as a long and multifunctional spacer arm covalently attached to an aminated surface capable of immobilizing DNA oligonucleotides. The immobilization occurred only via the amino introduced in the 3‘ end of the probe, with no implication of the DNA bases in the immobilization, ensuring that the full length of the probe is available for hybridization. These plates having immobilized oligonucleotide probes are able to hybridize complementary DNA target molecules. The tailor-made hetero-functional aldehyde-aspartic-dextran together with the chemical blocking of the remaining primary amino groups on the support using acetic anhydride avoid the nonspecific adsorption of DNA on the surface of the plates. Using these activated plates, (studying the effect of the probe concentration, temperature, and time of the plate activation on the achieved signal), thus, the covalent immobilization of the aminated DNA probe was optimized, and the sensitivity obtained was similar to that achieved using commercial biotin-streptavidin systems. The new DNA plates are stable under very drastic experimental conditions (90% formamide, at 100 °C for 30 min or in 100 mM NaOH).
Editor: Washington, DC: American Chemical Society
Idioma: Inglês

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Directed Covalent Immobilization of Aminated DNA Probes on Aminated Plates

Fuentes, Manuel ; Mateo, Cesar ; García, Lucia ; Tercero, Juan C ; Guisán, José M ; Fernández-Lafuente, Roberto

Washington, DC: American Chemical Society

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