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题名: Electrophoretic behavior of plasmid DNA in the presence of various intercalating dyes
作者: Dang, Fuquan;  Li, Wenhao;  Zhang, Lihua;  Jabasini, Mohammad;  Ishida, Tatsuhiro;  Kiwada, Hiroshi;  Kaji, Noritada;  Tokeshi, Manabu;  Baba, Yoshinobu
通讯作者: Yoshinobu Baba
关键词: plasmid DNA ;  supercoiled DNA ;  intercalating dyes ;  microchip ;  capillary electrophoresis ;  transfection
刊名: JOURNAL OF CHROMATOGRAPHY A
发表日期: 2006-06-23
DOI: 10.1016/j.chroma.2006.03.120
卷: 1118, 期:2, 页:218-225
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 3;3
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: In the present study, the electrophoretic behavior of linear, supercoiled and nicked circular plasmid DNA in the presence of various intercalating dyes was characterized using pGL3 plasmid DNA as a model. The enzymatic digestion of pGL3 plasmid DNA with HindIIIwas monitored by capillary electrophoresis coupled with laser-induced fluorescence detection (CE-LIF). Nicked circular plasmid DNA was found to be relatively sensitive to enzymes, and was almost digested into the linear conformer after 10-min incubation, indicating that nicked circular plasmid DNA has little chance of targeting and entering the cell nucleus. Partly digested plasmid DNA containing only linear and supercoiled conformers can be used as a standard to confirm the migration order of plasmid DNA. In methylcellulose (MC) solution with YO-PRO-1 or YOYO-1, linear plasmid DNA eluted first, followed by supercoiled and nicked plasmid DNA, and nicked plasmid DNA eluted as a broad peak. With SYBR Green 1, nicked plasmid DNA eluted first as three sharp peaks, followed by linear and supercoiled plasmid DNA. The nuclear plasmid DNA from two transfected cell fines was successfully analyzed using the present procedure. Similar results were obtained with an analysis time of seconds using microchip electrophoresis with laser-induced fluorescence detection (mu-CE-LIF). To our knowledge, these results represent the first reported analysis of nuclear plasmid DNA from transfection cells by CE-LIF or mu-CE-LIF without pre-preparation, suggesting that the present procedure is a promising alternative method for evaluating transfection efficiency of DNA delivery systems. (c) 2006 Elsevier B.V. All rights reserved.
关键词[WOS]: CAPILLARY ARRAY ELECTROPHORESIS ;  FLUORESCENCE DETECTION ;  GEL-ELECTROPHORESIS ;  DELIVERY-SYSTEMS ;  GENE DELIVERY ;  COPY NUMBER ;  SEPARATION ;  LIPOSOMES ;  MICROCHIP ;  GRADIENT
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000238311200007
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/98309
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.Nagoya Univ, Grad Sch Engn, Dept Appl Chem, Chikusa Ku, Nagoya, Aichi 4648603, Japan
2.Natl Inst Adv Ind Sci & Technol, AIST, Hlth Technol Res Ctr, Takamatsu, Kagawa 7610395, Japan
3.Univ Tokushima, Grad Sch Pharmaceut Sci, Tokushima 7708505, Japan
4.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian, Peoples R China

Recommended Citation:
Dang, Fuquan,Li, Wenhao,Zhang, Lihua,et al. Electrophoretic behavior of plasmid DNA in the presence of various intercalating dyes[J]. JOURNAL OF CHROMATOGRAPHY A,2006,1118(2):218-225.
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