Summary
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.
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Acknowledgments
We thank Jean-François Allemand for introduction to the combing method and members of our groups for helpful discussions.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Escudé, C., Géron-Landre, B., Crut, A., Desbiolles, P. (2009). Multicolor Detection of Combed DNA Molecules Using Quantum Dots. In: Foote, R., Lee, J. (eds) Micro and Nano Technologies in Bioanalysis. Methods in Molecular Biology™, vol 544. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-483-4_23
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DOI: https://doi.org/10.1007/978-1-59745-483-4_23
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