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. 2008 Nov 29;37(2):e10. doi: 10.1093/nar/gkn965

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© 2008 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Measurement of the unbinding forces between the 30-mer DNA probe on the AFM tip and the 802-base cRNA immobilized on a glass slide. (A) A schematic drawing of the experimental setup. The 802-base cRNA of the Pax6 mRNA containing the sequence complementary to the DNA probe sequence (red color, arrow) was immobilized on a glass slide. (B) A typical force–distance curve for the interaction between the DNA probe and cRNA. Forces were recorded at a measurement rate of 0.54 μm s⁻¹. (C) The histogram of unbinding forces derived from the force–distance curves of the interaction between the DNA probe and the cRNA. The histogram was obtained from 870 cycles of approach and retraction. The frequency of detection for unbinding events during the retraction process was 73% in this example. The Gaussian fitting gave the mean value of 41 ± 1 pN. The statistical error was estimated by , where σ is the width of the distribution of the N rupture events in the histogram. (D) The histograms of the unbinding forces between the DNA probe and antisense cRNA. The 802-base antisense RNA of Pax6 mRNA was synthesized in vitro and immobilized on a glass slide. The histogram was obtained from 657 cycles of approach and retraction.

Inline graphic — Measurement of the unbinding forces between the 30-mer DNA probe on the AFM tip and the 802-base cRNA immobilized on a glass slide. (A) A schematic drawing of the experimental setup. The 802-base cRNA of the Pax6 mRNA containing the sequence complementary to the DNA probe sequence (red color, arrow) was immobilized on a glass slide. (B) A typical force–distance curve for the interaction between the DNA probe and cRNA. Forces were recorded at a measurement rate of 0.54 μm s⁻¹. (C) The histogram of unbinding forces derived from the force–distance curves of the interaction between the DNA probe and the cRNA. The histogram was obtained from 870 cycles of approach and retraction. The frequency of detection for unbinding events during the retraction process was 73% in this example. The Gaussian fitting gave the mean value of 41 ± 1 pN. The statistical error was estimated by , where σ is the width of the distribution of the N rupture events in the histogram. (D) The histograms of the unbinding forces between the DNA probe and antisense cRNA. The 802-base antisense RNA of Pax6 mRNA was synthesized in vitro and immobilized on a glass slide. The histogram was obtained from 657 cycles of approach and retraction.