1. Marcaje no Enzimático y Detección no Radioactiva de ADN.
- Author
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Pérez-Bello, Dannelys, López-Dobarganes, Liliam, Riveron-Rojas, Ana Maria, Higginson-Clarke, David, Zayas, Mirta, Diaz, Nelson, Gonzalez, Ramón, Sablón Carrazana, Marquiza, Perez, Rafaela, and Rodriguez-Tanty, Chryslaine
- Subjects
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NUCLEIC acid probes , *DNA probes , *HYDROXYAPATITE , *CHROMATOGRAPHIC analysis , *MONOCLONAL antibodies - Abstract
Various enzymatic and chemical methods for the preparation of non-radioactive nucleic acid probes have been developed. Bisulfite-catalyzed transamination and acylation reactions into DNA are employed, as a chemical method, to obtain labeled DNA probes. If the reactions are carried out under DNA denaturalization condition, the labelling could be increased. Likewise, the label detection could be increased by means of the use of an appropriated spacer arm and also, by performing the label detection in single-stranded DNA. In this work, the procedure for obtainment labelled DNA probes, by chemical synthesis, using the p-bromobenzoyl radical as label was carried out. We developed a method for separating single-stranded DNA (s.s.DNA) from double stranded DNA, through Hydroxylapatite (HA3) chromatography. The sensitivity in the label detection was evaluated by chromogenic dot immunobinding assays using an anti-BLC monoclonal antibody (5H11E5), previously obtained. [ABSTRACT FROM AUTHOR]
- Published
- 2005