Center for Microfluidics and Medical Diagnostics, Department of Chemical and Biomolecular Engineering, University of Notre Dame, Notre Dame, IN 46556, USA.
Electrophoresis. 2010 Jan;31(4):666-71. doi: 10.1002/elps.200900473.
Oligonucleotides of varying surface coverage are functionalized onto the surface of 100 nm silica particles and the corresponding hybridization reaction with target ssDNA is studied using dielectrophoresis (DEP). The measured DEP cross-over frequency (cof) is found to be sensitive to the oligonucleotide surface conformation. Zeta potential and particle size measurements suggest that at low oligo surface concentrations, non-specific binding of oligo to the particle surface prevents efficient hybridization. At high surface coverage, steric hindrance due to the fully stretched, tightly packed oligo conformation prevents diffusion of DNA molecules to the particle surface. The optimum surface coverage exists at intermediate coverage where the particle is found to be the least electrically conductive, and hence exhibits the lowest measured cof. A simple DEP cof measurement hence allows one to determine the optimal oligo surface coverage for increased hybridization efficiency and detection sensitivity.
不同表面覆盖率的寡核苷酸被功能化到 100nm 二氧化硅颗粒的表面上,并使用介电泳(DEP)研究与靶 ssDNA 的相应杂交反应。所测量的 DEP 交叉频率(cof)对寡核苷酸表面构象敏感。zeta 电位和颗粒尺寸测量表明,在低寡核苷酸表面浓度下,寡核苷酸与颗粒表面的非特异性结合会阻止有效杂交。在高表面覆盖率下,由于完全伸展、紧密堆积的寡核苷酸构象而导致的空间位阻会阻止 DNA 分子扩散到颗粒表面。在最佳表面覆盖率下,颗粒的导电性最低,因此测量的 cof 也最低。因此,简单的 DEP cof 测量可以确定最佳的寡核苷酸表面覆盖率,以提高杂交效率和检测灵敏度。