High-Performance Genetic Analysis on Microfabricated Capillary Array Electrophoresis Plastic Chips Fabricated by Injection Molding
| Autor: | Osamu Tabata, Ashraf A. Ewis, Yoshihisa Yamaoka, Masaya Kurokawa, Yasuo Shinohara, Yoshinobu Baba, Lihua Zhang, Fuquan Dang, Mitsuru Ishikawa, Shouji Shinohara |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Capillary action
Analytical chemistry Fluorescence spectrometry Substrate (printing) Molding (process) Polymerase Chain Reaction Chemistry Techniques Analytical Analytical Chemistry law.invention Electrophoresis Microchip Capillary electrophoresis law Humans Deoxyribonucleases Type II Site-Specific Laser-induced fluorescence Miniaturization Polymorphism Genetic Pulmonary Surfactant-Associated Protein B business.industry Chemistry Electrophoresis Capillary Image intensifier DNA Equipment Design Globins Electrophoresis Mutation Optoelectronics business |
| Zdroj: | Analytical Chemistry. 77:2140-2146 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac0485031 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|