Department of Physics, University of Central Florida, Orlando, FL, 32816-2385, USA.
Sci Rep. 2021 May 7;11(1):9799. doi: 10.1038/s41598-021-89017-6.
The potential of a double nanopore system to determine DNA barcodes has been demonstrated experimentally. By carrying out Brownian dynamics simulation on a coarse-grained model DNA with protein tag (barcodes) at known locations along the chain backbone, we demonstrate that due to large variation of velocities of the chain segments between the tags, it is inevitable to under/overestimate the genetic lengths from the experimental current blockade and time of flight data. We demonstrate that it is the tension propagation along the chain's backbone that governs the motion of the entire chain and is the key element to explain the non uniformity and disparate velocities of the tags and DNA monomers under translocation that introduce errors in measurement of the length segments between protein tags. Using simulation data we further demonstrate that it is important to consider the dynamics of the entire chain and suggest methods to accurately decipher barcodes. We introduce and validate an interpolation scheme using simulation data for a broad distribution of tag separations and suggest how to implement the scheme experimentally.
双纳米孔系统用于确定 DNA 条码的潜力已经在实验中得到证实。通过对具有已知位置的蛋白质标签(条码)的粗粒化模型 DNA 进行布朗动力学模拟,我们证明由于标签之间链段速度的巨大差异,不可避免地会低估/高估实验电流阻塞和飞行时间数据中的遗传长度。我们证明,沿链骨架传播的张力控制着整个链的运动,是解释标签和 DNA 单体在易位过程中的非均匀性和不同速度的关键因素,这会导致在测量蛋白质标签之间的长度片段时产生误差。我们使用模拟数据进一步证明,考虑整个链的动力学非常重要,并提出了准确破译条码的方法。我们引入并验证了一种使用模拟数据的插值方案,该方案适用于广泛的标签分离分布,并提出了如何在实验中实施该方案的方法。