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 Scopus citations

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 - Nov 25 2003

Keywords

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

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry

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