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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)
KeywordChip Electrophoresis Integrated Microfluidic System Reaction-ce Microchip Restriction Digestion
Source PublicationELECTROPHORESIS
2008-12-01
DOI10.1002/elps.200800490
Volume29Issue:24Pages:4956-4963
Indexed BySCI
SubtypeArticle
Department18
Funding Project1807
Contribution Rank1;1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordPOLYMERASE-CHAIN-REACTION ; CAPILLARY ELECTROPHORESIS MICROSYSTEM ; NONSPECIFIC PROTEIN ADSORPTION ; MICROCHIP ELECTROPHORESIS ; SURFACE MODIFICATION ; POLY(DIMETHYLSILOXANE) ; MICRODEVICE ; SEPARATION ; PCR ; HYBRIDIZATION
AbstractA 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.
Language英语
URL查看原文
WOS IDWOS:000262565700018
Citation statistics
Cited Times:12[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/101591
Collection中国科学院大连化学物理研究所
Corresponding AuthorLin BC(林炳承)
Affiliation1.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
GB/T 7714
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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