FuGENE 6 transfection reagent: the gentle power

被引:46
作者
Jacobsen, LB [1 ]
Calvin, SA [1 ]
Colvin, KE [1 ]
Wright, M [1 ]
机构
[1] Roche Diagnost Corp, Roche Appl Sci, Indianapolis, IN 46250 USA
关键词
FuGENE 6 Transfection reagent; high throughput screening; antibiotic sensitivity; lipofection;
D O I
10.1016/j.ymeth.2003.11.002
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
FuGENE 6 Transfection Reagent has been commercially available since 1997. Since that time, its popularity has increased due to its ease of use, minimal to no cytotoxicity, and the high level of transfection in many different cell lines. FuGENE 6 Transfection Reagent is gentle on the cells. Adherent cells can be trypsinized and transfected by the DNA:FuGENE 6 reagent complex prior to plating, making it a strong candidate for high throughput applications. Additionally, low cell numbers can be transfected in 96-well plates. As with most reagents, the complex formation step is critical and special handling is required because the reagent is supplied in 80% ethanol. For example, contact with plastic must be avoided as inhibitors of transfection can leach from some plastics. We investigated parameters that have been reported to affect the transfection efficiency including the use of common antibiotics, passage level of the cells, and length of time for complex formation. These parameters are often cell line dependent and can be optimized to increase transfection efficiency for a specific cell line. (C) 2003 Elsevier Inc. All rights reserved.
引用
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页码:104 / 112
页数:9
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