Electrophoretic behavior of plasmid DNA in the presence of various intercalating dyes

	Electrophoretic behavior of plasmid DNA in the presence of various intercalating dyes
	Dang, Fuquan ; Li, Wenhao ; Zhang, Lihua ; Jabasini, Mohammad ; Ishida, Tatsuhiro ; Kiwada, Hiroshi ; Kaji, Noritada ; Tokeshi, Manabu ; Baba, Yoshinobu
刊名	journal of chromatography a
	2006-06-23
卷号	1118 期号:2 页码:218-225
关键词	plasmid DNA supercoiled DNA intercalating dyes microchip capillary electrophoresis transfection
通讯作者	yoshinobu baba
产权排序	3;3
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.
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]	capillary array electrophoresis ; fluorescence detection ; gel-electrophoresis ; delivery-systems ; gene delivery ; copy number ; separation ; liposomes ; microchip ; gradient
收录类别	SCI
原文出处	true
语种	英语
WOS记录号	WOS:000238311200007
公开日期	2010-11-30
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/98309]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
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
推荐引用方式 GB/T 7714	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.
APA	Dang, Fuquan.,Li, Wenhao.,Zhang, Lihua.,Jabasini, Mohammad.,Ishida, Tatsuhiro.,...&Baba, Yoshinobu.(2006).Electrophoretic behavior of plasmid DNA in the presence of various intercalating dyes.journal of chromatography a,1118(2),218-225.
MLA	Dang, Fuquan,et al."Electrophoretic behavior of plasmid DNA in the presence of various intercalating dyes".journal of chromatography a 1118.2(2006):218-225.