Imaging of single extended DNA molecules on flat (aminopropyl)triethoxysilane-mica by atomic force microscopy

Langmuir

Volumn 12, Issue 7, 1996, Pages

  Hu, J ^a,b,c   Wang, M ^a,d   Weier, H U G ^a,d   Frantz, P ^a,b   Kolbe, W ^a,e   Ogletree, D F ^a,b   Salmeron, M ^a,b  

^a UNIVERSITY OF CALIFORNIA   (United States)

^b Materials Sciences Division

^c INSTITUTE OF GEOLOGY AND GEOPHYSICS   (China)

^d Life Sciences Division

^e Electrical Engineering Division

Author keywords

[No Author keywords available]

Indexed keywords

EID: 5244348938     PISSN: 07437463     EISSN: None     Source Type: Journal    
DOI: None     Document Type: Article

References (15)

1. Binnig, G.; Quate, C. F.; Gerber, C. Phys. Rev. Lett. 1986, 56, 930.
2. Rugar, D.; Hansma, P. Phys. Today 1990, 43, 23.
3. Hansma, P. K.; Cleveland, J. P.; Radmacher, M.; Walters, D. A.; Hillner, P. E.; Bezanilla, M.; Fritz, M.; Vie, D.; Hansma, H. G. Appl. Phys. Lett. 1994, 64, 1738.
4. Hansma, H. G.; Bezanilla, M.; Zenhausern, F.; Adrian, M.; Sinsheimer, R. L. Nucleic Acids Res. 1993, 21, 505.
5. Bezanilla, M.; Drake, B.; Nudler, E.; Kashlev, M.; Hansma, P. K.; Hansma, H. G. Biophys. J. 1994, 67, 2454.
6. Bezanilla, M.; Bustamante, C.; Hansma, H. G. Scanning Microsc. 1994, 7, 1145.
7. Guthold, M.; Bezanilla, M.; Jenkins, B.; Hansma, H. G.; Bustamante, C. Proc. Natl. Acad. Sci. U.S.A. 1994, 91, 12927.
8. Lyubchenko, Y. L.; Gall, A. A.; Shlyyakhtenko, L. S.; Harrington, R. E.; Jacobs, B. L.; Oden, P. I.; Lindsay, S. M. J. Biomol. Struct. Dyn. 1992, 10, 589.
9. Lyubchenko, Y. L.; Shlyyakhtenko, L. S.; Harrington, R. E.; Oden, P. I.; Lindsay, S. M. Proc. Natl. Acad. Sci. U.S.A. 1993, 90, 2137.
10. Bezanilla, M.; Manne, S.; Laney, D. E.; Lyubchenco, Y. L.; Hansma, H. G. Langmuir 1996, 11, 655.
11. Bensimon, A.; Simon, A.; Chiffaudel, A.; Croquette, V.; Heslot, F.; Bensimon, D. Science 1994, 265, 2096.
12. Weier, H.-U. G.; Wang, M.; Mullikin, J. C.; Zhu, Y.; Cheng, J.-F.; Greulich, K. M.; Bensimon, A.; Gray, J. W. Hum. Mol. Genet. 1995, 4, 1903.
13. Detailed results are to be published elsewhere. Briefly, an aqueous solution of APS, 1% by volume, is used here. The container must be very clean. A freshly cleaved mica sheet is immersed into the APS solution for several minutes. APS forms a thin, disordered monolayer ∼5-8 Å thick on the mica surface. The surface of APS-mica prepared in this way is very flat, with a roughness of ∼3 Å over several micrometers. The charge state of the amine group of APS on mica is dependent upon the pH of the solution.
14. Kolbe, W. F.; Ogletree, D. F.; Salmeron, M. B. Ultramicroscopy 1992, 42-44, 1113.
15. DNA was deposited and extended onto APS-mica substrate and denatured by heating at 95 °C for 2 min. The sample was then overlaid with the solution containing Klenow Fragment of DNA polymeraee I, random primers from GIBCO/LTI's BioPrime DNA Labeling System, and a deoxynucleoside triphosphate mixture containing biotin-dCTP. After incubating at 37 °C overnight, slides were washed in 2x SSC and incubated with avidin-FITC for 30 min at 20 °C. Finally, the slides were washed in two changes of 2x SSC and the sample was observed under a fluorescence microscope. The biotin-labeled DNA which was synthesized during extension of the random primers was visible in the microscope as long and continuous green DNA strands. Control samples treated in the same way, except that the Klenow Fragment of DNA Polymerase I was omitted from the buffer, did not produce DNA strands labeled and visible in the fluorescence microscope. These results indicated that DNA synthesis and replication processes occurred along the extended and adsorbed DNA strands. Detailed results will be published elsewhere.