Experimental Biophysics and Applied Nanoscience, Faculty of Physics, Bielefeld University, Bielefeld, Germany.
Anal Chem. 2010 Sep 1;82(17):7141-9. doi: 10.1021/ac1005475.
Dielectrophoresis is a convenient tool for controlled manipulation of DNA with numerous applications, including DNA trapping, stretching, and separation. However, the mechanisms behind the dielectrophoretic properties of DNA are still under debate, and the role of conformation has not been addressed yet. Here, we quantify dielectrophoretic effects on DNA by determining its polarizability from microfluidic single molecule trapping experiments. We systematically study different DNA configurations (linear and supercoiled, 6-164 kbp) and demonstrate that the polarizability strongly depends on the specific conformation and size of the DNA molecules. The connection to its spatial extension is established by measuring diffusion coefficients and from that the radii of gyration; details about the spatial DNA structure are obtained from atomic force microscopy images. For linear and supercoiled DNA fragments, we found a power-law scaling for the polarizabilities and the diffusion coefficients. Our results imply a scaling of the polarizability with the radius of gyration, alpha approximately Rg0.9+/-0.1 and alpha approximately Rg1.6+/-0.2 for linear and supercoiled DNA, respectively. As an application, we demonstrate the separation of DNA topoisomers based on their dielectrophoretic properties, achieving baseline resolution within 210 s. Purified DNA samples of specific configuration may be of great importance for DNA nanoassembly or future DNA vaccines.
介电泳是一种方便的工具,可用于控制 DNA 的操作,具有多种应用,包括 DNA 捕获、拉伸和分离。然而,DNA 介电泳性质背后的机制仍存在争议,构象的作用尚未得到解决。在这里,我们通过从微流控单分子捕获实验中确定其介电常数来量化 DNA 的介电泳效应。我们系统地研究了不同的 DNA 构象(线性和超螺旋,6-164 kbp),并证明介电常数强烈依赖于 DNA 分子的特定构象和大小。通过测量扩散系数并由此确定转动半径,建立了与空间延伸的联系;通过原子力显微镜图像获得有关空间 DNA 结构的详细信息。对于线性和超螺旋 DNA 片段,我们发现介电常数和扩散系数呈幂律关系。我们的结果表明,介电常数与转动半径的标度关系为,alpha approximately Rg0.9+/-0.1 和 alpha approximately Rg1.6+/-0.2,分别适用于线性和超螺旋 DNA。作为一种应用,我们证明了基于其介电泳性质的 DNA 拓扑异构体的分离,在 210 秒内实现了基线分辨率。特定构象的纯化 DNA 样本可能对 DNA 纳米组装或未来的 DNA 疫苗非常重要。