Burnham Daniel R, De Vlaminck Iwijn, Henighan Thomas, Dekker Cees
Delft University of Technology, Kavli Institute of Nanoscience, Department of Bionanoscience, Delft, The Netherlands.
PLoS One. 2014 Sep 29;9(9):e108271. doi: 10.1371/journal.pone.0108271. eCollection 2014.
Measurements in magnetic tweezers rely upon precise determination of the position of a magnetic microsphere. Fluctuations in the position due to Brownian motion allows calculation of the applied force, enabling deduction of the force-extension response function for a single DNA molecule that is attached to the microsphere. The standard approach relies upon using the mean of position fluctuations, which is valid when the microsphere axial position fluctuations obey a normal distribution. However, here we demonstrate that nearby surfaces and the non-linear elasticity of DNA can skew the distribution. Through experiment and simulations, we show that such a skewing leads to inaccurate position measurements which significantly affect the extracted DNA extension and mechanical properties, leading to up to two-fold errors in measured DNA persistence length. We develop a simple, robust and easily implemented method to correct for such mismeasurements.
磁镊测量依赖于对磁性微球位置的精确测定。由于布朗运动导致的位置波动可用于计算所施加的力,从而能够推导附着在微球上的单个DNA分子的力-伸长响应函数。标准方法依赖于使用位置波动的平均值,当微球轴向位置波动服从正态分布时,该方法是有效的。然而,我们在此证明,附近的表面和DNA的非线性弹性会使分布产生偏差。通过实验和模拟,我们表明这种偏差会导致位置测量不准确,从而显著影响提取的DNA伸长和力学性能,导致测量的DNA持久长度出现高达两倍的误差。我们开发了一种简单、稳健且易于实施的方法来校正这种测量误差。