Duke T A, Austin R H, Cox E C, Chan S S
Physics Department, Princeton University, NJ 08544, USA.
Electrophoresis. 1996 Jun;17(6):1075-9. doi: 10.1002/elps.1150170616.
Transverse pulsed-field electrophoresis of DNA has been conducted in a silicon array engineered by optical lithography and the motion of individual molecules observed by fluorescence microscopy. In strong fields, the molecules can be maintained in highly stretched, linear conformations. When the field is switched through an obtuse angle, they head off in the new direction led by what was formerly their tail end. This backtracking gives rise to fractionation that is linear with molecular weight. A simple prescription exists for choosing the field parameters to obtain a particular range of separation. Since the molecular motions are much more uniform than those that occur in a gel, it is anticipated that the arrays will permit more efficient fractionation than traditional pulsed-field gel electrophoresis. Arrays suitably scaled down in size may be useful for pulsed-field sequencing.
已在通过光刻技术制造的硅阵列中进行了DNA的横向脉冲场电泳，并通过荧光显微镜观察了单个分子的运动。在强电场中，分子可以保持高度拉伸的线性构象。当电场以钝角切换时，它们会在由原来的尾端引导的新方向上前进。这种回溯导致了与分子量成线性关系的分离。存在一种简单的方法来选择场参数以获得特定的分离范围。由于分子运动比凝胶中的运动更加均匀，预计该阵列将比传统的脉冲场凝胶电泳实现更高效的分离。尺寸适当缩小的阵列可能对脉冲场测序有用。
