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

被引:52
作者
Dang, FQ
Tabata, O
Kurokawa, M
Ewis, AA
Zhang, LH
Yamaoka, Y
Shinohara, S
Shinohara, Y
Ishikawa, M
Baba, Y
机构
[1] AIST, Simgle Mol Bioanal Lab, Takamatsu, Kagawa 7610395, Japan
[2] Kyoto Univ, Grad Sch Engn, Dept Mech Engn, Sakyo Ku, Kyoto 6068501, Japan
[3] Starlite Co Ltd, Kusatsu 5203004, Japan
[4] Univ Tokushima, Grad Sch Pharmaceut Sci, CREST, JST,Dept Mol & Phamaceut Biotechnol, Tokushima 7708505, Japan
[5] Japan Sci & Technol Corp, Kawaguchi, Saitama 3320012, Japan
[6] Nagoya Univ, Grad Sch Engn, Dept Appl Chem, Chikusa Ku, Nagoya, Aichi 4648603, Japan
关键词
D O I
10.1021/ac0485031
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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-mu m width, 50-mu m depth, and a 100-mu m 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 phi X174 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.
引用
收藏
页码:2140 / 2146
页数:7
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