Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (263K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Géron-Landre, B.
Right arrow Articles by Escudé, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Géron-Landre, B.
Right arrow Articles by Escudé, C.
Related Collections
Right arrow Nucleic acid amplification
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2003, Vol. 31, No. 20 e125
© 2003 Oxford University Press

Sequence-specific fluorescent labeling of double-stranded DNA observed at the single molecule level

Bénédicte Géron-Landre, Thibaut Roulon, Pierre Desbiolles1 and Christophe Escudé*

Laboratoire de Biophysique, Muséum National d’Histoire Naturelle, INSERM U565, CNRS UMR8646, 43, rue Cuvier, 75231 Paris Cedex 05, France and 1 Laboratoire Kastler Brossel, Ecole Normale Supérieure, 24, rue Lhomond, 75005 Paris, France

*To whom correspondence should be addressed. Tel: +33 1 40793774; Fax: +33 1 40793705; Email: escude{at}mnhn.fr
Present address:
Thibaut Roulon, ANOSYS Inc., 1014 Hamilton Court, Menlo Park, CA 94025, USA
This paper is dedicated to the memory of Professor Claude Hélène. We are grateful to him for initiating this project, for sharing with us his numerous ideas and for transmitting his passion for research in the field of nucleic acids.

Fluorescent labeling of a short sequence of double-stranded DNA (dsDNA) was achieved by ligating a labeled dsDNA fragment to a stem–loop triplex forming oligonucleotide (TFO). After the TFO has wound around the target sequence by ligand-induced triple helix formation, its extremities hybridize to each other, leaving a dangling single-stranded sequence, which is then ligated to a fluorescent dsDNA fragment using T4 DNA ligase. A non-repeated 15 bp sequence present on lambda DNA was labeled and visualized by fluorescence microscopy after DNA combing. The label was found to be attached at a specific position located at 4.2 ± 0.5 kb from one end of the molecule, in agreement with the location of the target sequence for triple helix formation (4.4 kb from one end). In addition, an alternative combing process was noticed in which a DNA molecule becomes attached to the combing slide from the label rather than from one of its ends. The method described herein provides a new tool for the detection of very short sequences of dsDNA and offers various perspectives in the micromanipulation of single DNA molecules.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
M. Mendez-Lago, J. Wild, S. L. Whitehead, A. Tracey, B. de Pablos, J. Rogers, W. Szybalski, and A. Villasante
Novel sequencing strategy for repetitive DNA in a Drosophila BAC clone reveals that the centromeric region of the Y chromosome evolved from a telomere
Nucleic Acids Res., April 1, 2009; 37(7): 2264 - 2273.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Duca, P. Vekhoff, K. Oussedik, L. Halby, and P. B. Arimondo
The triple helix: 50 years later, the outcome
Nucleic Acids Res., September 1, 2008; 36(16): 5123 - 5138.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
H. Kuhn and M. D. Frank-Kamenetskii
Labeling of unique sequences in double-stranded DNA at sites of vicinal nicks generated by nicking endonucleases
Nucleic Acids Res., April 1, 2008; 36(7): e40 - e40.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
A. Crut, B. Geron-Landre, I. Bonnet, S. Bonneau, P. Desbiolles, and C. Escude
Detection of single DNA molecules by multicolor quantum-dot end-labeling
Nucleic Acids Res., June 20, 2005; 33(11): e98 - e98.
[Abstract] [Full Text] [PDF]

Home page
 Genome ResHome page
Y. Shigemori, H. Haruta, T. Okada, and M. Oishi
Marking of specific sequences in double-stranded DNA molecules--SNP detection and direct observation
Genome Res., December 1, 2004; 14(12): 2478 - 2485.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.