TY  - JOUR
A1  - Stopper, Helga
A1  - Zimmermann, U.
A1  - Neil, G. A.
T1  - Increased efficiency of transfection of murine hybridoma cells with DNA by electropermeabilization
N2  - Dispase-treated murine hybridoma cells (SP2/0-Ag14) were transfected with the G418 resistance gene bearing plasmid pSV2-neo by electropermeabilization with a high degree of efficiency. The cells were subjected to intermittent multiple high-voltage short duration (5 p.s) DC pulses at intervals of 1 min in a weakly conducting medium followed by selection in G418-containing medium. The transfection medium, temperature, pulse duration, and voltage were empirically determined by preliminary electropermeabilization experiments. Increasing the number of pulses resulted in a higher percentage of transfected cells, but a decrease in the number of viable cells, with the optimal transfectant yield resulting when five pulses of 10 kV jcm were administered. This method allows the rapid and efficient injection of DNA into mammalian cells, and permits the rapid production of stable, drug resistant hybridoma celllines for use in subsequent fusion experiments.
KW  - Toxikologie
KW  - DNA transfection
KW  - Electropermeabilization
KW  - Eukaryotic cell
KW  - Hybridoma
KW  - Drug resistance
Y1  - 1988
UR  - https://opus.bibliothek.uni-wuerzburg.de/frontdoor/index/index/docId/6032
UR  - https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-63488
ER  -