Advanced transfection with Lipofectamine 2000 reagent: primary neurons, siRNA, and high-throughput applications

被引:456
作者
Dalby, B
Cates, S
Harris, A
Ohki, EC
Tilkins, ML
Price, PJ
Ciccarone, VC
机构
[1] Invitrogen Corp, Res & Dev, Carlsbad, CA 92008 USA
[2] Invitrogen Corp, GIBCO Cell Culture Div, Grand Isl, NY 14072 USA
[3] MacroGenics Inc, Rockville, MD 20850 USA
关键词
D O I
10.1016/j.ymeth.2003.11.023
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Lipofectamine 2000 is a cationic liposome based reagent that provides high transfection efficiency and high levels of transgene expression in a range of mammalian cell types in vitro using a simple protocol. Optimum transfection efficiency and subsequent cell viability depend on a number of experimental variables such as cell density, liposome and DNA concentrations, liposome-DNA complexing time, and the presence or absence of media components such as antibiotics and serum. The importance of these factors in Lipofectamine 2000 mediated transfection will be discussed together with some specific applications: transfection of primary neurons, high throughput transfection, and delivery of small interfering RNAs. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:95 / 103
页数:9
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