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Microfluidic device for integrated restriction digestion reaction and resulting DNA fragment analysis
Xie, Hua1,2; Li, Bowei1,2; Zhong, Runtao1,2; Qin, Jianhua1; Zhu, Yisheng3; Lin, Bingcheng1; Lin BC(林炳承)
关键词Chip Electrophoresis Integrated Microfluidic System Reaction-ce Microchip Restriction Digestion
刊名ELECTROPHORESIS
2008-12-01
DOI10.1002/elps.200800490
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
关键词[WOS]POLYMERASE-CHAIN-REACTION ; CAPILLARY ELECTROPHORESIS MICROSYSTEM ; NONSPECIFIC PROTEIN ADSORPTION ; MICROCHIP ELECTROPHORESIS ; SURFACE MODIFICATION ; POLY(DIMETHYLSILOXANE) ; MICRODEVICE ; SEPARATION ; PCR ; HYBRIDIZATION
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.
语种英语
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WOS记录号WOS:000262565700018
引用统计
被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/101591
专题中国科学院大连化学物理研究所
通讯作者Lin BC(林炳承)
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
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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.
APA Xie, Hua.,Li, Bowei.,Zhong, Runtao.,Qin, Jianhua.,Zhu, Yisheng.,...&林炳承.(2008).Microfluidic device for integrated restriction digestion reaction and resulting DNA fragment analysis.ELECTROPHORESIS,29(24),4956-4963.
MLA Xie, Hua,et al."Microfluidic device for integrated restriction digestion reaction and resulting DNA fragment analysis".ELECTROPHORESIS 29.24(2008):4956-4963.
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