用于毛细管凝胶电泳的微加工聚合物芯片。

Microfabricated polymer chip for capillary gel electrophoresis.

作者信息

Hong J W, Hosokawa K, Fujii T, Seki M, Endo I

机构信息

Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Tokyo 113-8656, Japan.

出版信息

Biotechnol Prog. 2001 Sep-Oct;17(5):958-62. doi: 10.1021/bp010075m.

DOI:10.1021/bp010075m

PMID:11587590

Abstract

A polymer (PDMS: poly(dimethylsiloxane)) microchip for capillary gel electrophoresis that can separate different sizes of DNA molecules in a small experimental scale is presented. This microchip can be easily produced by a simple PDMS molding method against a microfabricated master without the use of elaborate bonding processes. This PDMS microchip could be used as a single use device unlike conventional microchips made of glass, quartz or silicon. The capillary channel on the chip was partially filled with agarose gel that can enhance separation resolution of different sizes of DNA molecules and can shorten the channel length required for the separation of the sample compared to capillary electrophoresis in free-flow or polymer solution format. We discuss the optimal conditions for the gel preparation that could be used in the microchannel. DNA molecules were successfully driven by an electric field and separated to form bands in the range of 100 bp to 1 kbp in a 2.0% agarose-filled microchannel with 8 mm of effective separation length.

摘要

本文介绍了一种用于毛细管凝胶电泳的聚合物（聚二甲基硅氧烷，PDMS）微芯片，它能够在小规模实验中分离不同大小的DNA分子。这种微芯片可以通过简单的PDMS成型方法，以微加工母版为模板轻松制备，无需使用复杂的键合工艺。与由玻璃、石英或硅制成的传统微芯片不同，这种PDMS微芯片可以作为一次性装置使用。芯片上的毛细管通道部分填充有琼脂糖凝胶，与自由流动或聚合物溶液形式的毛细管电泳相比，琼脂糖凝胶可以提高不同大小DNA分子的分离分辨率，并缩短样品分离所需的通道长度。我们讨论了可用于微通道的凝胶制备的最佳条件。在有效分离长度为8 mm、填充2.0%琼脂糖的微通道中，DNA分子在电场驱动下成功分离，形成了100 bp至1 kbp范围内的条带。