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Improved transfection efficiency of cultured human cells
Author(s) -
Nikcevic Gordana,
KovacevicGrujicic Natasa,
Stevanovic Milena
Publication year - 2003
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/s1065-6995(03)00143-4
Subject(s) - lipofectamine , transfection , hela , plasmid , microbiology and biotechnology , cell culture , reporter gene , cytotoxicity , chemistry , dna , biology , recombinant dna , gene , in vitro , vector (molecular biology) , biochemistry , gene expression , genetics
We simultaneously tested the transfection efficiency of NT2/D1 and HeLa cells with Lipofectamine™ (Life Technologies) and Effectene™ (Qiagen) transfection reagents using the pCH110 eukaryotic assay vector, which contains the lac Z reporter gene. Under our culture conditions for NT2/D1 and HeLa cells, Effectene™ transfection efficiency could be augmented by simply increasing the amount of plasmid DNA 1.5–3 times above the recommended concentration without any visible cytotoxicity. With the Lipofectamine™ reagent, optimal transfection efficiency was obtained for both cell lines within the recommended concentrations, but at the top of the range. The results indicate that optimization of the transfection process should include plasmid DNA concentrations above the levels suggested by the manufacturers, in order to accomplish the highest transfection efficiency.