Application of on-chip capillary electrophoresis to cell-free preparation of recombinant DNA.
An on-chip capillary electrophoresis-based DNA collection was applied to the isolation of target DNA species from a DNA mixture generated by a polymerase chain reaction (PCR), whose starting material was a ligation mixture of an insert and an expression vector. The collected DNA was then amplified by PCR and properly worked as template DNA in a coupled cell-free transcription/translation system. These results demonstrateed that total operation in standard genetic engineering can be performed in a cell-free condition.
Nojima
Takahiko
T
Center for International Research on MicroMechatronics, Institute of Industrial Science, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan. nojima@iis.u-tokyo.ac.jp
Kaneda
Shohei
S
Fujii
Teruo
T
D002474:Cell-Free System; D004274:DNA, Recombinant / Q000302:isolation & purification*; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D013698:Templates, Genetic; D014158:Transcription, Genetic
D004274:DNA, Recombinant