Non-enzymatic in vitro DNA labeling and label immunoquantification

David Higginson Clarke, Dannelys Pérez-Bello, Ana María Riverón-Rojas, Guillermo Jiménez-Alemán, Mirta Zayas Acosta, Rafaela Pérez Perera, Marquiza Sablón Carrazana, Nelson Díaz Miranda, Chryslaine Rodríguez-Tanty, Esteban Gutiérrez Calzado

Research output: Contribution to journalArticle

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Abstract

In the present work, a label immunoquantification procedure was developed in order to determine the number of markers introduced into DNA. A non-enzymatic, in vitro labeling method for introducing the p-bromobenzoyl radical (label), through transamination and acylation reactions of the cytidine nucleotides in calf thymus DNA, was carried out. Three spacer arms with different lengths were used for separating the label from the nucleotide and three labeled DNA were obtained. Anti-p-bromobenzoyl chicken IgY polyclonal antibodies were obtained. The antibodies detected the label, into three-labeled DNA, with different sensitivities, in relation to spacer arm length used. About 3-11 labels per 4 × 106 bases into thermally denatured DNA were immunoquantified.

Original languageEnglish (US)
Pages (from-to)1-15
Number of pages15
JournalPreparative Biochemistry and Biotechnology
Volume35
Issue number1
DOIs
StatePublished - Feb 2 2005

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Keywords

  • Bisulfite-catalyzed transamination reaction
  • Chicken IgY polyclonal antibodies
  • In vitro non-radioactive DNA labeling
  • Label immunoquantification

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry

Cite this

Clarke, D. H., Pérez-Bello, D., Riverón-Rojas, A. M., Jiménez-Alemán, G., Acosta, M. Z., Perera, R. P., Carrazana, M. S., Miranda, N. D., Rodríguez-Tanty, C., & Calzado, E. G. (2005). Non-enzymatic in vitro DNA labeling and label immunoquantification. Preparative Biochemistry and Biotechnology, 35(1), 1-15. https://doi.org/10.1081/PB-200041428