Comparative Study of Three Methods for Non-radioactive in Vivo DNA Labeling in Escherichia coli Using Nucleoside Analogs

Dannelys Pérez-Bello, Viana Manrique Suárez, Celso Pérez Bolaños, Judith Miranda López, Gabriel Pérez Pérez, Lilia López-Canovas, David Higginson-Clarke, Chryslaine Rodríguez-Tanty, Ana María Riverón

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In the present work, a comparative study of 5-FdUrd, thy-, and metabolic in vivo labeling methods for plasmid and chromosomal DNA in E. coli DH5α cells was performed in order to achieve the best thymidine substitution method by 5-BrdUrd. According to the colorimetric immunoenzymatic results, we found that the minimal detectable labeled DNA (MDLD) was 312pg with the 5-FdUrd and thy methods for 5-BrdUrd labeled plasmid DNA. 5-BrdUrd replaced about 96% of the total thymidine by 5-FdUrd methods; for the thy- and metabolic labeling methods, the MDLD value was 1,25ng for denatured 5-BrdUrd chromosomal DNA. Pyrimidine nucleoside analogues were also evaluated as immunochemical markers for their in vivo introduction into DNA.

Original languageEnglish (US)
Pages (from-to)253-268
Number of pages16
JournalPreparative Biochemistry and Biotechnology
Volume33
Issue number4
DOIs
StatePublished - 2003
Externally publishedYes

Fingerprint

Nucleosides
Labeling
Escherichia coli
DNA
Thymidine
Plasmids
Pyrimidine Nucleosides
Substitution reactions

Keywords

  • 5-BrdUrd
  • In vivo labeling methods
  • Non-radioactive probe
  • Pyrimidine nucleoside analogues

ASJC Scopus subject areas

  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Biotechnology

Cite this

Pérez-Bello, D., Suárez, V. M., Bolaños, C. P., López, J. M., Pérez, G. P., López-Canovas, L., ... Riverón, A. M. (2003). Comparative Study of Three Methods for Non-radioactive in Vivo DNA Labeling in Escherichia coli Using Nucleoside Analogs. Preparative Biochemistry and Biotechnology, 33(4), 253-268. https://doi.org/10.1081/PB-120025369

Comparative Study of Three Methods for Non-radioactive in Vivo DNA Labeling in Escherichia coli Using Nucleoside Analogs. / Pérez-Bello, Dannelys; Suárez, Viana Manrique; Bolaños, Celso Pérez; López, Judith Miranda; Pérez, Gabriel Pérez; López-Canovas, Lilia; Higginson-Clarke, David; Rodríguez-Tanty, Chryslaine; Riverón, Ana María.

In: Preparative Biochemistry and Biotechnology, Vol. 33, No. 4, 2003, p. 253-268.

Research output: Contribution to journalArticle

Pérez-Bello, D, Suárez, VM, Bolaños, CP, López, JM, Pérez, GP, López-Canovas, L, Higginson-Clarke, D, Rodríguez-Tanty, C & Riverón, AM 2003, 'Comparative Study of Three Methods for Non-radioactive in Vivo DNA Labeling in Escherichia coli Using Nucleoside Analogs', Preparative Biochemistry and Biotechnology, vol. 33, no. 4, pp. 253-268. https://doi.org/10.1081/PB-120025369
Pérez-Bello, Dannelys ; Suárez, Viana Manrique ; Bolaños, Celso Pérez ; López, Judith Miranda ; Pérez, Gabriel Pérez ; López-Canovas, Lilia ; Higginson-Clarke, David ; Rodríguez-Tanty, Chryslaine ; Riverón, Ana María. / Comparative Study of Three Methods for Non-radioactive in Vivo DNA Labeling in Escherichia coli Using Nucleoside Analogs. In: Preparative Biochemistry and Biotechnology. 2003 ; Vol. 33, No. 4. pp. 253-268.
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