title: Multicolor detection of combed DNA molecules using quantum dots.
abstract: DNA combing is a useful strategy for manipulating single DNA molecules and has a wide range of applications in genetics, single molecule studies, and nanobiotechnology. Visualization of combed DNA molecules is usually performed by using DNA binding organic dyes. Such dyes are not suitable in all circumstances, especially because of their photoreactivity. We have developed a method for the detection of combed DNA molecules by fluorescence microscopy that avoids the use of DNA-staining agents and does not perturb the structure of the DNA molecule. Biotin- and/or digoxigenin-modified DNA fragments are covalently linked at both ends of a DNA molecule via sequence-specific hybridization and subsequent ligation. After the modified DNA molecules have been combed on a polystyrene-coated surface, their ends are visualized by multicolor fluorescence microscopy using conjugated quantum dots.
authors.affiliation: Régulations et Dynamique des Génomes, INSERM U565, Muséum National d'Histoire Naturelle, Paris Cedex 05, France. escude@mnhn.fr
authors.forename: Christophe
authors.initials: C
authors.lastname: Escudé
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Géron-Landre
authors.forename: Aurélien
authors.initials: A
authors.lastname: Crut
authors.forename: Pierre
authors.initials: P
authors.lastname: Desbiolles
mesh_terms: D001583:Benzoxazoles; D001710:Biotin; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D004076:Digoxigenin; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence; D045663:Quantum Dots; D011806:Quinolinium Compounds
chemical_list: D001583:Benzoxazoles; D005456:Fluorescent Dyes; D011806:Quinolinium Compounds; C075296:1,1'-((4,4,7,7-tetramethyl)-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-oxazole)-2-methylidene)quinolinium; D001710:Biotin; D004247:DNA; D004076:Digoxigenin