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对通过注塑成型制造的微型毛细管阵列电泳塑料芯片进行高性能基因分析。

High-performance genetic analysis on microfabricated capillary array electrophoresis plastic chips fabricated by injection molding.

作者信息

Dang Fuquan, Tabata Osamu, Kurokawa Masaya, Ewis Ashraf A, Zhang Lihua, Yamaoka Yoshihisa, Shinohara Shouji, Shinohara Yasuo, Ishikawa Mitsuru, Baba Yoshinobu

机构信息

Single-Molecule Bioanalysis Laboratory, National Institute of Advanced Industrial Science and Technology (AIST), Hayashi-cho 2217-14, Takamatsu 761-0395, Japan.

出版信息

Anal Chem. 2005 Apr 1;77(7):2140-6. doi: 10.1021/ac0485031.

Abstract

We have developed a novel technique for mass production of microfabricated capillary array electrophoresis (mu-CAE) plastic chips for high-speed, high-throughput genetic analysis. The mu-CAE chips, containing 10 individual separation channels of 50-microm width, 50-microm depth, and a 100-microm lane-to-lane spacing at the detection region and a sacrificial channel network, were fabricated on a poly(methyl methacrylate) substrate by injection molding and then bonded manually using a pressure-sensitive sealing tape within several seconds at room temperature. The conditions for injection molding and bonding were carefully characterized to yield mu-CAE chips with well-defined channel and injection structures. A CCD camera equipped with an image intensifier was used to monitor simultaneously the separation in a 10-channel array with laser-induced fluorescence detection. High-performance electrophoretic separations of phiX174 HaeIII DNA restriction fragments and PCR products related to the human beta-globin gene and SP-B gene (the surfactant protein B) have been demonstrated on mu-CAE plastic chips using a methylcellulose sieving matrix in individual channels. The current work demonstrated greatly simplified the fabrication process as well as a detection scheme for mu-CAE chips and will bring the low-cost mass production and application of mu-CAE plastic chips for genetic analysis.

摘要

我们已经开发出一种用于大规模生产微制造毛细管阵列电泳(mu-CAE)塑料芯片的新技术,用于高速、高通量基因分析。mu-CAE芯片包含10个独立的分离通道,通道宽度为50微米,深度为50微米,在检测区域的通道间间距为100微米,还有一个牺牲通道网络。该芯片通过注塑成型在聚甲基丙烯酸甲酯基板上制造,然后在室温下使用压敏密封胶带在几秒钟内手动键合。对注塑成型和键合条件进行了仔细表征,以生产出具有明确通道和进样结构的mu-CAE芯片。配备图像增强器的CCD相机用于通过激光诱导荧光检测同时监测10通道阵列中的分离情况。使用甲基纤维素筛分基质在单个通道中,已在mu-CAE塑料芯片上展示了phiX174 HaeIII DNA限制性片段以及与人β-珠蛋白基因和SP-B基因(表面活性剂蛋白B)相关的PCR产物的高效电泳分离。当前工作极大地简化了mu-CAE芯片的制造过程以及检测方案,并将实现mu-CAE塑料芯片用于基因分析的低成本大规模生产和应用。

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