An optimized electroporation protocol applicable to a wide range of cell lines.
A number of transfection methods for mammalian cells are available; however, many cell lines may appear resistant to efficient transfection, or at best, necessitate lengthy optimization procedures in recommended protocols. We describe here an electroporation protocol that yields highly efficient gene transfer (20%-100% of surviving cells) in all 19 cell lines tested so far, including lymphoid, myeloid, glial, fibroblast and COS cells. Unlike earlier electroporation protocols, adaptation of this protocol to a cell line of interest only requires the optimization of a single parameter, i.e., the voltage, and can be performed within a day. Furthermore, it is also superior to transfection protocols for other methods (calcium phosphate precipitation, lipofection, DEAE-dextran transfection and transferrinfection) in terms of reproducibility, economy and average transfection efficiency for a wide variety of cell lines.
Baum
C
C
Heinrich-Pette-Institut, Universität Hamburg, FRG.
Forster
P
P
Hegewisch-Becker
S
S
Harbers
K
K
D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis; D018274:Electroporation / Q000379:methods*; D005434:Flow Cytometry; D005697:Galactosides / Q000032:analysis; D000882:Haplorhini; D006801:Humans; D007211:Indoles / Q000032:analysis; D051379:Mice; D051381:Rats; D014162:Transfection / Q000379:methods*
D005697:Galactosides; D007211:Indoles; D015500:Chloramphenicol O-Acetyltransferase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside