title: A novel DNA selection and direct extraction process and its application in DNA recombination.
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.
authors.affiliation: Institute of Applied Mechanics, National Taiwan University, Taipei, Taiwan. abwang@spring.iam.ntu.edu.tw
authors.forename: An-Bang
authors.initials: AB
authors.lastname: Wang
authors.forename: Chia-Wei
authors.initials: CW
authors.lastname: Cheng
authors.forename: I-Chun
authors.initials: IC
authors.lastname: Lin
authors.forename: Fei-Yau
authors.initials: FY
authors.lastname: Lu
authors.forename: Huai-Jen
authors.initials: HJ
authors.lastname: Tsai
authors.forename: Chiu-Chun
authors.initials: CC
authors.lastname: Lin
authors.forename: Chun-Hui
authors.initials: CH
authors.lastname: Yang
authors.forename: Po-Ting
authors.initials: PT
authors.lastname: Pan
authors.forename: Chen-Chi
authors.initials: CC
authors.lastname: Kuan
authors.forename: Yen-Chih
authors.initials: YC
authors.lastname: Chen
authors.forename: Yi-Wei
authors.initials: YW
authors.lastname: Lin
authors.forename: Chih-Ning
authors.initials: CN
authors.lastname: Chang
authors.forename: Yi-Hung
authors.initials: YH
authors.lastname: Wu
authors.forename: Tetuko
authors.initials: T
authors.lastname: Kurniawan
authors.forename: Chii-Wann
authors.initials: CW
authors.lastname: Lin
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Wo
authors.forename: Lin-Chi
authors.initials: LC
authors.lastname: Chen
mesh_terms: D004247:DNA / Q000032:analysis*; D004586:Electrophoresis / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D044085:Microfluidics / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011995:Recombination, Genetic / Q000235:genetics*
chemical_list: D004247:DNA