title: Analysis of specific gene by integration of isothermal amplification and electrophoresis on poly(methyl methacrylate) microchips.
abstract: We have successfully achieved the integration of isothermal amplification and the subsequent analysis of specific gene fragments on poly(methyl methacrylate) microchips. In our experiments, loop-mediated isothermal amplification, which can offer higher specificity and efficiency than PCR, has been performed at a constant temperature (65 degrees C). After amplification, products could be either examined by the integrated microchip-based electrophoresis or directly observed by naked eye with SYBR Green I added into the reaction solution. By such an integrated microsystem, the amplification and the subsequent analysis of prostate-specific antigen gene with template concentration at 23 fg/microL could be finished within 15 min, which demonstrates its advantages of high specificity, good reproducibility, and fast speed in gene detection.
authors.affiliation: Department of Molecular and Pharmaceutical Biotechnology, Graduate School of Pharmaceutical Sciences, The University of Tokushima, 21st Century COE, CREST, JST, Shomachi, Tokushima 770-8505, Japan.
authors.forename: Yukari
authors.initials: Y
authors.lastname: Hataoka
authors.forename: Lihua
authors.initials: L
authors.lastname: Zhang
authors.forename: Yasuyohi
authors.initials: Y
authors.lastname: Mori
authors.forename: Norihiro
authors.initials: N
authors.lastname: Tomita
authors.forename: Tsugunori
authors.initials: T
authors.lastname: Notomi
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Baba
mesh_terms: D004247:DNA / Q000032:analysis* / Q000235:genetics; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D005796:Genes / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D017430:Prostate-Specific Antigen / Q000235:genetics*; D012680:Sensitivity and Specificity; D013499:Surface Properties; D013696:Temperature
chemical_list: D004247:DNA; D019904:Polymethyl Methacrylate; D017430:Prostate-Specific Antigen