A novel DNA selection and direct extraction process and its application in DNA recombination |
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| Authors: | Wang An-Bang Cheng Chia-Wei Lin I-Chun Lu Fei-Yau Tsai Huai-Jen Lin Chiu-Chun Yang Chun-Hui Pan Po-Ting Kuan Chen-Chi Chen Yen-Chih Lin Yi-Wei Chang Chih-Ning Wu Yi-Hung Kurniawan Tetuko Lin Chii-Wann Wo Andrew M Chen Lin-Chi |
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| Affiliation: | Institute of Applied Mechanics, National Taiwan University, Taipei, Taiwan. abwang@spring.iam.ntu.edu.tw |
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| Abstract: | In the conventional bench-top approach, the DNA recombination process is time- and effort-consuming due to laborious procedures lasting from several hours to a day. A novel DNA selection and direct extraction process has been proposed, integrated and tested on chip. The integrative microfluidic chip can perform the whole procedure of DNA recombination, including DNA digestion, gel electrophoresis, DNA extraction and insert-vector ligation within 1?h. In this high-throughput design, the manual gel cutting was replaced by an automatic processing system that performed high-quality and high-recovery efficiency in DNA extraction process. With no need of gel-dissolving reagents and manipulation, the application of selection and direct extraction process could significantly eliminate the risks from UV and EtBr and also facilitate DNA recombination. Reliable output with high success rate of cloning has been achieved with a significant reduction in operational hazards, required materials, efforts and time. |
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