Abstract
DNA molecules were stretched on silanized mica surface with the molecular combing technique, and detected with fluorescence microscopy and atomic force microscopy. Meantime, DNA molecules were stretched with a modified dynamic molecular combing technique and studied with atomic force microscopy. The results indicate that, compared with the dynamic molecular combing technique, the modified dynamic molecular combing technique has advantages of less-sample demand and less contamination to sample; as compared with the molecular combing technique, it has better aligning effect and reproducibility. Combination of this kind of DNA molecular manipulating technique with the single DNA molecule detecting technique by atomic force microscopy and fluorescence microscopy will play an important role in the basic research of molecular dynamics and the application of gene research.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bensimon, A., Simon, A., Chiffaudel, A. et al., Alignment and sensitive detection of DNA by a moving interface, Science, 1994, 265: 2096.
Weier, H.U. G., Wang, M., Mullikin, J. C. et al., Quantitative DNA fiber mapping. Human Molecular Genetics, 1995, 4(10): 1903.
Michalet, X., Ekong, R., Fougerousse, F. et al., Dynamic molecular combing: stretching the whole human genome for high-resolution studies. Science, 1997,277: 1518.
Cai, W., Aburatani, H., Stanton, V. P. et al., Ordered restriction endonuclease maps of yeast artificial chromosomes created by optical mapping on surfaces. Proc. Natl. Acad. Sci., 1995,92: 5164.
Ouyang, Z. Q., Hu, J., Chen, S. F. et al., Molecular patterns by manipulating DNA molecules, J. Vac. Sci. Technol. B, 1997,15(4): 1385.
Zimmermann, R. M., Cox, E. C., DNA stretching on functionalized gold surfaces, Nucleic Acids Research, 1994, 22(3): 492.
Li, J. W., Bai, C. L., Wang, C. et al., A convenient method of aligning large DNA molecules on bare mica surfaces for atomic force microscopy. Nucleic Acids Research, 1998,26(20): 4785.
Hu, J., Wang, M., Weier, H.-U. G. et al., Imaging of single extended DNA molecules on flat (Aminopropyl) triethoxysilane-mica by atomic force microscopy, Langmuir, 1996, 12(7): 1697.
Li, M. Q., Scanning probe microscopy (STM/AFM) and applications in biology, Applied Physics A, 1999,68: 1.
Jing, J., Reed, J., Huang, J. et al., Automated high resolution optical mapping using arrayed, fluid-fixed DNA molecules, Proc. Natl. Acad. Sci. USA, 1998,95: 8046.
Bustamante, C., Direct observation and manipulation of single DNA molecules using fluorescence microscopy, Annu. Rev. Biophys. Biochem., 1991,20: 415.
Glazer, A. N., Rye, H. S., Stable dye-DNA intercalation complexes as reagents for high-sensitivity fluorescence, Nature, 1992,359: 859.
Gurrieri, G., Wetl, K. S., Johnson, I. D. et al., Direct visualization of individual DNA molecules by fluorescence microscopy: characterization of the factors affecting signal/background and optimization of imaging conditions using YOYO, Analytical Biochemistry, 1997,249: 44.
Florijn, R. J., Bonden, L. A. J., Vrolijk, H. et al., High-resolution DNA fiber-FISH for genomic DNA mapping and color bar-coding of large genes, Human Molecular Genetics, 1995,4(5): 831.
Xu, X. M., Ikai, A., Measurement of mechanical properties of human chromosomes using atomic force microscopy, Cetlular Engineering, 1997,2(2): 75.
Xu, X. M., Ikai, A., Recovery and amplification of plasmid DNA with atomic force microscopy and the polymerase chain reaction, Analytica Chimica Acta, 1998,361: 1.
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Zhang, Y., Chen, S., Ouyang, Z. et al. Stretching and imaging studies of single DNA molecules. Chin. Sci. Bull. 45, 1365–1368 (2000). https://doi.org/10.1007/BF02886238
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02886238