Detection of single DNA molecules by multicolor quantum-dot end-labeling - Archive ouverte HAL Access content directly
Journal Articles Nucleic Acids Research Year : 2005

Detection of single DNA molecules by multicolor quantum-dot end-labeling

Aurelien Crut
Bénédicte Géron-Landre
  • Function : Author
Stéphane Bonneau
  • Function : Author


Observation of DNA-protein interactions by single molecule fluorescence microscopy is usually performed by using fluorescent DNA binding agents. However, such dyes have been shown to induce cleavage of the DNA molecule and perturb its interactions with proteins. A new method for the detection of surface-attached DNA molecules by fluorescence microscopy is introduced in this paper. Biotin-and/or digoxigenin-modified DNA fragments are covalently linked at both extremities of a DNA molecule via sequence-specific hybridization and ligation. After the modified DNA molecules have been stretched on a glass surface, their ends are visualized by mul-ticolor fluorescence microscopy using conjugated quantum dots (QD). We demonstrate that under carefully selected conditions, the position and orientation of individual DNA molecules can be inferred with good efficiency from the QD fluores-cence signals alone. This is achieved by selecting QD pairs that have the distance and direction expected for the combed DNA molecules. Direct observation of single DNA molecules in the absence of DNA staining agent opens new possibilities in the fundamental study of DNA-protein interactions. This work also documents new possibilities regarding the use of QD for nucleic acid detection and analysis.
Fichier principal
Vignette du fichier
Crut_NAR_2005_QDotDNA.pdf (749.19 Ko) Télécharger le fichier
Origin Publication funded by an institution

Dates and versions

hal-02454251 , version 1 (24-01-2020)




Aurelien Crut, Bénédicte Géron-Landre, Isabelle Bonnet, Stéphane Bonneau, Pierre Desbiolles, et al.. Detection of single DNA molecules by multicolor quantum-dot end-labeling. Nucleic Acids Research, 2005, 33 (11), pp.e98-e98. ⟨10.1093/nar/gni097⟩. ⟨hal-02454251⟩
48 View
44 Download



Gmail Mastodon Facebook X LinkedIn More