transient transfection efficiency
Posted 23 May 2009 - 03:32 PM
I am transfecting Drosophila cells with a plasmid that has a GFP gene under a constitutive promoter. I see only a few percent of the cells are GFP positive with either method. Is this typical? I was hoping it would be a little more efficient.
I am especially interested in the situations with the Drosophila cells but comments on mammalian cells would be helpful, too. Thanks.
Posted 24 May 2009 - 04:38 PM
Posted 26 May 2009 - 04:53 AM
I've only used mammalian cells - typical transfection efficiency would be about 70-90% depending a bit on the cell line and the plasmid being transfected. Some cell types are much harder to transfect than others, you could have one of those.
Choice of transfection reagent is critical to transient transfection in different cell types. Some cell types may show 90% transfection with lipofectamine 2000 (for example) but only 20% with Effectene, while other cells show the reverse response. You should search the literature to determine the optimal reagent for your cell type. Also, check sites like Invitrogen, Roche, biorad, and others for their reagents. Oftentimes, they will list different cell types for which their reagents have been tested and the determined efficiencies. You can also ask for a free sample to test on your cell types. You should aim for >70% efficiency if you want to make any real conclusions. If none of these options work, try electroporation or viral infection.
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