DNA ELECTROPHORESIS IN MICROLITHOGRAPHIC ARRAYS
DNA ELECTROPHORESIS IN MICROLITHOGRAPHIC ARRAYS
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DOI：
10.1038/358600a0
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发表时间：
1992-08-13
期刊：
影响因子：
64.8
通讯作者：
AUSTIN, RH
中科院分区：
文献类型：
--
作者：
VOLKMUTH, WD;AUSTIN, RH
我们利用光学微光刻技术在二氧化硅中制造了有覆盖层的准二维障碍通道。在此我们报道利用落射荧光显微镜对限制在阵列中的大DNA分子的电泳和长度分级的观察结果。基于德热纳的研究成果的简单爬行理论预测，在低电场下，长度\(L\)远大于持续长度\(p\)的聚合物的电泳迁移率与\(L\)成反比（参考文献2）。但是在足够强的电场下，聚合物链在基质中的伸长会导致电泳迁移率与长度无关（参考文献4，5）。施加适当定时的脉冲电场可恢复凝胶对长分子的分级能力（参考文献6），但是脉冲场电泳的操作流程是半经验性的，因为复杂且尚未充分了解的凝胶基质在分级过程中起着关键作用。通过微光刻构建的障碍阵列，由于其低维度、小体积以及极其可重复的形貌，将使我们有可能了解大聚合物分子在复杂但特征明确的拓扑结构中的运动和分级情况。
WE have used optical microlithography to fabricate capped quasi-two-dimensional obstacle courses in SiO2. We report here observations using epifluorescence microscopy of the electrophoresis and length fractionation of large DNA molecules confined in arrays. Simple reptation theory, based on the work of deGennes1, predicts that at low electric fields the electrophoretic mobility of a polymer of length L much greater than the persistence length p scales inversely with L (ref. 2). But elongation of the coil in the matrix at sufficiently strong electric fields3 results in a length-independent electrophoretic mobility4,5. The application of suitably timed pulsed electric fields restores the fractionating power of gels for long molecules6 but the protocols of pulsed-field electrophoresis are semi-empirical because the complex and ill-understood gel matrix plays a critical role in fractionation. Microlithographically constructed obstacle arrays, with their low dimensionality, small volume and extremely reproducible topography, will make it possible to understand the motion and fractionation of large polymer molecules in complex but well characterized topologies.