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题名: Microfluidic device for integrated restriction digestion reaction and resulting DNA fragment analysis
作者: Xie, Hua1, 2;  Li, Bowei1, 2;  Zhong, Runtao1, 2;  Qin, Jianhua1;  Zhu, Yisheng3;  Lin, Bingcheng1
通讯作者: 林炳承
关键词: Chip electrophoresis ;  Integrated microfluidic system ;  Reaction-CE microchip ;  Restriction digestion
刊名: ELECTROPHORESIS
发表日期: 2008-12-01
DOI: 10.1002/elps.200800490
卷: 29, 期:24, 页:4956-4963
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1807
产权排名: 1;1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: A microfluidic system combining tempera tu re-controlled reactor, analyte delivery, chip electrophoresis (CE) separation, and fluorescence detection was developed, in which the heaters, resistance temperature detectors, enzymatic reactors, CE channels, and pneumatic valves/pumps were integrated onto a single glass-PDMS chip. The microdevice was used to perform the digestion reaction, followed by on-line electrophoresis separation and detection of the resulting fragments with endonuclease BamHI and FokI as models. Pneumatic valves/pumps served not only for isolating the reaction region from the separation medium to prevent contamination, but also for delivering and quantitatively diluting the fluid from the reaction chamber to the CE section. Thus enzymatic reaction and electrophoresis separation could be insulated and connected as needed. A dynamic coating procedure with the use of PVP and mannitol was firstly adopted for glass-PDMS hybrid chip-based DNA separations, leading to an improved separation efficiency with reproducible migration time and theoretical plates. The expected 263- and 287-bp digestion products of BamHI and FokI were definitely verified by the size-based electrophoretic separation and detection. The whole integrated reaction-CE system can be manipulated in a simple manner with good reproducibility, which is expected to be applied in other on-line analysis of various biochemical reactions.
关键词[WOS]: POLYMERASE-CHAIN-REACTION ;  CAPILLARY ELECTROPHORESIS MICROSYSTEM ;  NONSPECIFIC PROTEIN ADSORPTION ;  MICROCHIP ELECTROPHORESIS ;  SURFACE MODIFICATION ;  POLY(DIMETHYLSILOXANE) ;  MICRODEVICE ;  SEPARATION ;  PCR ;  HYBRIDIZATION
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000262565700018
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/101591
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
3.Shanghai Jiao Tong Univ, Dept Biomed Engn, Shanghai 200030, Peoples R China

Recommended Citation:
Xie, Hua,Li, Bowei,Zhong, Runtao,et al. Microfluidic device for integrated restriction digestion reaction and resulting DNA fragment analysis[J]. ELECTROPHORESIS,2008,29(24):4956-4963.
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