Stable transfection cells selection

Stable transfection of cells with selectable vectors... 

The targeting construct will be electroporated into ES cells. After selection for stable transfectants expressing a specific antibiotic resistance (here neomycin resistance, neo), homologous recombinants will be identified and injected into blastocysts. Transfer of injected blastocysts into specially conditioned ( pseudopregnant") foster mothers results in chimeric mice that consist... 

Electroporation condition Selection condition MSX [pM] Number of stable transfectants per 5x10 cells electroporated... 

An assessment of the transfection efficiency of human skin fibroblasts, compared to transfection of hamster lung fibroblasts and Human HeLa cells, indicated that the protocols we used are not suitable to obtain stable transfected skin fibroblasts. We concluded this from the fact that no stable transfected colonies of human skin fibroblasts were obtained, regardless of being CPTl-defective or being from a patient with a disorder unrelated to P-oxidation. Only transient expression with the Lipofectamine method was successful. We found an efficiency of 0.0-0.2 % of the cells which survived transfection without selection. However, compared to the efficiencies of transfection in hamster lung fibroblasts (5-10% in a parallel experiment), this still was low. Therefore, for long-term apphcations it is clear that different procedures should be used than the ones we applied here. 

Introduction Vector Design Inducible Systems Cell Lines Transfection Methods Transient vs. Stable Transfection Selectable Markers Single Cell Cloning Methods... 

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