Integrated capillary electrophoresis on flexible silicone microdevices: Analysis of DNA restriction fragments and detection of single DNA molecules on microchips
Integrated capillary electrophoresis on flexible silicone microdevices: Analysis of DNA restriction fragments and detection of single DNA molecules on microchips
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DOI：
10.1021/ac9703919
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发表时间：
1997-09-01
影响因子：
7.4
通讯作者：
Ehrat, M
中科院分区：
文献类型：
--
作者：
Effenhauser, CS;Bruin, GJM;Ehrat, M
用于集成毛细管电泳系统的微芯片是通过在微加工母模上模制聚（二甲基硅氧烷）（PDMS）有机硅弹性体来生产的。 PDMS 装置在干净的平面上具有良好的粘附性，可以简单且廉价地生成密封微通道网络，从而消除了复杂粘合程序的限制，通过快速分离用嵌入染料 YOYO-1 标记的 phi X-174/HaeIII DNA 限制性片段和荧光标记的肽，证明了装置的性能。 603 碱基对片段的质量检测限类似于 2 fg。可以使用简单的激光诱导荧光检测装置检测以 10:1 的碱基对与染料比率插入 YOYO-1 的单个 lambda-DNA 分子，在 PDMS 微通道中的电泳控制传质条件下实现了高单分子检测效率 (>50%)。
Microchips for integrated capillary electrophoresis systems were produced by molding a poly(dimethylsiloxane) (PDMS) silicone elastomer against a microfabricated master. The good adhesion of the PDMS devices on clean planar surfaces allows for a simple and inexpensive generation of networks of sealed microchannels, thus removing the constraints of elaborate bonding procedures, The performance of the devices is demonstrated with both fast separations of phi X-174/HaeIII DNA restriction fragments labelled with the intercalating dye YOYO-1 and fluorescently labelled peptides, Detection limits in the order of a few zeptomoles (10(-21) mel) have been achieved for each injected DNA fragment, corresponding to a mass detection limit of similar to 2 fg for the 603 base pair fragment. Single lambda-DNA molecules intercalated with YOYO-1 at a base pair-to-dye ratio of 10:1 could be detected with an uncomplicated laser-induced fluorescence detection setup, High single-molecule detection efficiency (>50%) was achieved under electrophoretically controlled mass transport conditions in PDMS microchannels.