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使用微机电仪器的高泳道密度平板凝胶电泳

High lane density slab-gel electrophoresis using micromachined instrumentation.

作者信息

Papautsky I, Mohanty S, Weiss R, Frazier A B

机构信息

Department of Electrical and Computer Engineering and Computer Science, University of Cincinnati, OH 45221, USA.

出版信息

Electrophoresis. 2001 Oct;22(18):3908-15. doi: 10.1002/1522-2683(200110)22:18<3908::AID-ELPS3908>3.0.CO;2-L.

Abstract

In this paper, micromachined pipette arrays (MPAs) and microcombs were studied as a means of enabling high lane density gel electrophoresis. The MPA provide a miniaturized format to interface sub-microliter volumes of samples between macroscale sample preparation formats and microscale biochemical analysis systems. The microcombs provide a means of creating sample loading wells in the gel material on the same center-to-center spacing as the MPAs. Together, the two micromachined instruments provide an alternative to current combs and pipetting technologies used for creating sample loading wells and sample delivery in gel electrophoresis systems. Using three designs for the microcomb-MPA pair, center-to-center spacings of 1.0 mm, 500 microm, and 250 microm are studied. The results demonstrate an approximate 10-fold increase in lane density and a 10-fold reduction in sample size from 5 microL to 500 pL. As a result, the number of theoretical plates has increased 2.5-fold, while system resolution has increased 1.5-fold over the conventional agarose gel systems. An examination of changes in resolution across the width of individual separation lanes in both systems revealed dependence in the case of the conventional gels and no dependence for the gels loaded with the micromachined instrumentation.

摘要

在本文中,对微加工移液器阵列(MPA)和微梳进行了研究,将其作为实现高泳道密度凝胶电泳的一种手段。MPA提供了一种小型化形式,用于在宏观样品制备形式和微观生化分析系统之间连接亚微升体积的样品。微梳提供了一种在凝胶材料中创建样品加样孔的方法,其中心距与MPA相同。这两种微加工仪器共同为目前用于在凝胶电泳系统中创建样品加样孔和样品输送的梳子和移液技术提供了一种替代方案。使用微梳-MPA对的三种设计,研究了1.0毫米、500微米和250微米的中心距。结果表明,泳道密度增加了约10倍,样品大小从5微升减少到500皮升,减少了10倍。因此,理论塔板数增加了2.5倍,而系统分辨率比传统琼脂糖凝胶系统提高了1.5倍。对两个系统中单个分离泳道宽度上分辨率变化的检查表明,传统凝胶存在依赖性,而使用微加工仪器加载的凝胶则不存在依赖性。

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