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 | |
| State | Published - 2003 |
| Externally published | Yes |
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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
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 journal › Article
}
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
Y1 - 2003
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 -