High-performance genetic analysis on microfabricated capillary array electrophoresis plastic chips fabricated by injection molding.
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.
Dang
Fuquan
F
Single-Molecule Bioanalysis Laboratory, National Institute of Advanced Industrial Science and Technology (AIST), Hayashi-cho 2217-14, Takamatsu 761-0395, Japan. fuquandang@aist.go.jp
Tabata
Osamu
O
Kurokawa
Masaya
M
Ewis
Ashraf A
AA
Zhang
Lihua
L
Yamaoka
Yoshihisa
Y
Shinohara
Shouji
S
Shinohara
Yasuo
Y
Ishikawa
Mitsuru
M
Baba
Yoshinobu
Y
D002623:Chemistry Techniques, Analytical / Q000379:methods; D004247:DNA / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods*; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D005914:Globins / Q000235:genetics; D006801:Humans; D008904:Miniaturization; D009154:Mutation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D037701:Pulmonary Surfactant-Associated Protein B / Q000235:genetics
D037701:Pulmonary Surfactant-Associated Protein B; D005914:Globins; D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific; C024403:GGCC-specific type II deoxyribonucleases