A novel DNA selection and direct extraction process and its application in DNA recombination
Journal
Electrophoresis
Journal Volume
32
Journal Issue
3-4
Pages
423-430
Date Issued
2011
Author(s)
Cheng C.-W.
Lin I.-C.
Lu F.-Y.
Tsai H.-J.
Lin C.-C.
Yang C.-H.
Pan P.-T.
Kuan C.-C.
Chen Y.-C.
Lin Y.-W.
Chang C.-N.
Wu Y.-H.
Kurniawan T.
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 1h. 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. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Subjects
Chip integration; DNA recombination; DNA selection and extraction; Microfluidics
Other Subjects
ethidium bromide; article; automation; DNA degradation; DNA determination; DNA extraction; DNA recombination; gel electrophoresis; microfluidics; risk management; ultraviolet radiation; DNA; Electrophoresis; Equipment Design; Equipment Failure Analysis; Microfluidic Analytical Techniques; Microfluidics; Oligonucleotide Array Sequence Analysis; Recombination, Genetic
Type
journal article