Vanderhoeven Johan, Pappaert Kris, Dutta Binita, Vanhummelen Paul, Baron Gino V, Desmet Gert
Vrije Universiteit Brussel, Department of Chemical Engineering, Brussels, Belgium.
Electrophoresis. 2004 Nov;25(21-22):3677-86. doi: 10.1002/elps.200406116.
The present study demonstrates that the best way to enhance DNA microarray assays, both in terms of analysis speed and in final spot intensity, is to dissolve the available molar amount of sample in the smallest possible buffer volume and to subsequently convect this solution continuously across the surface of the array. The presently proposed shear-driven flow system is pre-eminently suited for this task, as it allows to induce strongly enhanced lateral transport rates, independently of the degree of miniaturization of the hybridization chamber. This transport enhancement method, however, only increases the hybridization rate and not the final spot intensity, as neither can any of the other transport enhancement methods already proposed in literature. A series of experiments with synthetic single-stranded (ssDNA) samples and an accompanying mass balance analysis are presented to demonstrate these points.
本研究表明，要在分析速度和最终斑点强度方面增强DNA微阵列检测，最佳方法是将可用摩尔量的样品溶解在尽可能小的缓冲液体积中，然后使该溶液持续对流穿过阵列表面。目前提出的剪切驱动流系统非常适合这项任务，因为它能够独立于杂交室的小型化程度诱导大幅增强的横向传输速率。然而，这种传输增强方法仅提高杂交速率，而不会提高最终斑点强度，文献中已提出的其他传输增强方法也都做不到这一点。本文展示了一系列使用合成单链（ssDNA）样品的实验以及伴随的质量平衡分析来证明这些观点。
