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Elasticity of Short DNA Molecules: Theory and Experiment for Contour Lengths of 0.6–7 μm

机译:短DNA分子的弹性:0.6–7μm轮廓长度的理论和实验

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摘要

The wormlike chain (WLC) model currently provides the best description of double-stranded DNA elasticity for micron-sized molecules. This theory requires two intrinsic material parameters—the contour length L and the persistence length p. We measured and then analyzed the elasticity of double-stranded DNA as a function of L (632 nm–7.03 μm) using the classic solution to the WLC model. When the elasticity data were analyzed using this solution, the resulting fitted value for the persistence length pwlc depended on L; even for moderately long DNA molecules (L = 1300 nm), this apparent persistence length was 10% smaller than its limiting value for long DNA. Because p is a material parameter, and cannot depend on length, we sought a new solution to the WLC model, which we call the “finite wormlike chain (FWLC),” to account for effects not considered in the classic solution. Specifically we accounted for the finite chain length, the chain-end boundary conditions, and the bead rotational fluctuations inherent in optical trapping assays where beads are used to apply the force. After incorporating these corrections, we used our FWLC solution to generate force-extension curves, and then fit those curves with the classic WLC solution, as done in the standard experimental analysis. These results qualitatively reproduced the apparent dependence of pwlc on L seen in experimental data when analyzed with the classic WLC solution. Directly fitting experimental data to the FWLC solution reduces the apparent dependence of pfwlc on L by a factor of 3. Thus, the FWLC solution provides a significantly improved theoretical framework in which to analyze single-molecule experiments over a broad range of experimentally accessible DNA lengths, including both short (a few hundred nanometers in contour length) and very long (microns in contour length) molecules.
机译:蠕虫状链(WLC)模型目前提供了微米级分子的双链DNA弹性的最佳描述。该理论需要两个固有的材料参数-轮廓长度L和持久长度p。我们使用经典的WLC模型解决方案测量并分析了双链DNA的弹性随L(632 nm–7.03μm)的变化。当使用该解决方案分析弹性数据时,持久长度pwlc的最终拟合值取决于L;即使对于中等长度的DNA分子(L = 1300 nm),该表观持久性长度也比其对长DNA的极限值小10%。由于p是材料参数,并且不能取决于长度,因此我们寻求WLC模型的新解决方案,我们将其称为“有限蠕虫链(FWLC)”,以解决经典解决方案中未考虑的影响。具体来说,我们考虑了有限的链长,链端边界条件以及在使用珠子施加力的光学诱捕测定法中固有的珠子旋转波动。合并这些校正后,我们使用FWLC解决方案生成力-伸长曲线,然后将这些曲线与经典WLC解决方案进行拟合,如标准实验分析中所做的那样。当使用经典WLC解决方案进行分析时,这些结果从质上再现了pwlc对L的表观依赖性。直接将实验数据拟合到FWLC解决方案中,可将pfwlc对L的表观依赖性降低3倍。因此,FWLC解决方案提供了显着改进的理论框架,可在其中分析广泛的实验可访问的DNA长度上的单分子实验,包括短分子(轮廓长度几百纳米)和长分子(轮廓长度几微米)。

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