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

Anesthesiology

Research output: Contribution to journal › Article

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 - Nov 25 2003

Fingerprint

Keywords

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

ASJC Scopus subject areas

Biotechnology
Biochemistry

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

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, 25.11.2003, p. 253-268.

Research output: Contribution to journal › Article

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.

@article{7ac89cc25acd4ab0b9ca55260fbd9906,

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

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.",

keywords = "5-BrdUrd, In vivo labeling methods, Non-radioactive probe, Pyrimidine nucleoside analogues",

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}",

year = "2003",

month = nov

day = "25",

doi = "10.1081/PB-120025369",

language = "English (US)",

volume = "33",

pages = "253--268",

journal = "Preparative Biochemistry and Biotechnology",

issn = "1082-6068",

publisher = "Taylor and Francis Ltd.",

number = "4",

}

TY - JOUR

T1 - Comparative Study of Three Methods for Non-radioactive in Vivo DNA Labeling in Escherichia coli Using Nucleoside Analogs

AU - Pérez-Bello, Dannelys

AU - Suárez, Viana Manrique

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

PY - 2003/11/25

Y1 - 2003/11/25

N2 - 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.

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.

KW - 5-BrdUrd

KW - In vivo labeling methods

KW - Non-radioactive probe

KW - Pyrimidine nucleoside analogues

UR - http://www.scopus.com/inward/record.url?scp=0242490733&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0242490733&partnerID=8YFLogxK

U2 - 10.1081/PB-120025369

DO - 10.1081/PB-120025369

M3 - Article

C2 - 14606684

AN - SCOPUS:0242490733

VL - 33

SP - 253

EP - 268

JO - Preparative Biochemistry and Biotechnology

JF - Preparative Biochemistry and Biotechnology

SN - 1082-6068

IS - 4

ER -

Access to Document

10.1081/PB-120025369