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© Research
Publication : Molecular and cellular probes

5-Bromodeoxyuridine in vivo labelling of M13 DNA, and its use as a non-radioactive probe for hybridization experiments

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Molecular and cellular probes - 01 Mar 1987

Sakamoto H, Traincard F, Vo-Quang T, Ternynck T, Guesdon JL, Avrameas S

Link to Pubmed [PMID] – 3331168

Mol. Cell. Probes 1987 Mar;1(1):109-20

We describe the in vivo production of 5-bromodeoxyuridine- (5-BUdR) labelled M13 DNA by a thymine-requiring Escherichia coli strain. We show that the 5-BUdR-labelled M13 single-stranded DNA is not extruded into the culture medium, but accumulates inside the bacterial cells. On the basis of this observation, a procedure involving FPLC gel filtration already reported and used for the isolation of plasmid DNA has been adapted for the isolation of at least 90% pure 5-BUdR-labelled single-stranded DNA. An M13 probe, containing part of the Hepatitis B Virus (HBV) genome was constructed, and the corresponding 5-BUdR-labelled single-stranded DNA was used in hybridization experiments to detect homologous HBV target DNA. Picogram amounts (10(-19) moles) of the probe itself or the target DNA could be detected, by monoclonal anti-5-BUdR antibodies in an immunoenzymatic assay.

http://www.ncbi.nlm.nih.gov/pubmed/3331168