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

doi:10.1081/PB-120025369

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 journal › Article › peer-review

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 language	English (US)
Pages (from-to)	253-268
Number of pages	16
Journal	Preparative Biochemistry and Biotechnology
Volume	33
Issue number	4
DOIs	https://doi.org/10.1081/PB-120025369
State	Published - 2003
Externally published	Yes

Keywords

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

ASJC Scopus subject areas

Biotechnology
Biochemistry

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10.1081/PB-120025369

Cite this

Pérez-Bello, D., Suárez, V. M., Bolaños, C. P., López, J. M., Pérez, G. P., López-Canovas, L., Higginson-Clarke, D., Rodríguez-Tanty, C., & 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

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

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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.",

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author = "Dannelys P{\'e}rez-Bello and Su{\'a}rez, {Viana Manrique} and Bola{\~n}os, {Celso P{\'e}rez} and L{\'o}pez, {Judith Miranda} and P{\'e}rez, {Gabriel P{\'e}rez} and Lilia L{\'o}pez-Canovas and David Higginson-Clarke and Chryslaine Rodr{\'i}guez-Tanty and River{\'o}n, {Ana Mar{\'i}a}",

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AU - Pérez-Bello, Dannelys

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AU - Bolaños, Celso Pérez

AU - López, Judith Miranda

AU - Pérez, Gabriel Pérez

AU - López-Canovas, Lilia

AU - Higginson-Clarke, David

AU - Rodríguez-Tanty, Chryslaine

AU - Riverón, Ana María

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AB - 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.

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KW - Non-radioactive probe

KW - Pyrimidine nucleoside analogues

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

Abstract

Keywords

ASJC Scopus subject areas

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