Dokument-ID Dokumenttyp Verfasser/Autoren Herausgeber Haupttitel Abstract Auflage Verlagsort Verlag Erscheinungsjahr Seitenzahl Schriftenreihe Titel Schriftenreihe Bandzahl ISBN Quelle der Hochschulschrift Konferenzname Quelle:Titel Quelle:Jahrgang Quelle:Heftnummer Quelle:Erste Seite Quelle:Letzte Seite URN DOI Abteilungen OPUS4-6044 Wissenschaftlicher Artikel Stopper, Helga; Jones, H.; Zimmermann, U. Large scale transfection of mouse L-cells by electropermeabilization Mouse L-cells were transfected by electropenneabilization using the selectable plasmid pSV2-neo which confers resistance to G-418 (Geneticin). 1be DNA concentration used was 1 l'gfml, the field strength was 10 kV fcm, the duration of the pulse was S ~s. Transfeetion yield was optimal at a temperature of 4°C when using a time in between consecutive pulses of 1 minute compared to shorter (of the order of seoonds) or Ionger (3 minutes) time intervals. A more detailed study of the relationship between the number of pulses applied (up to 10) and transfection yield showed it to be almost linear in this range at 4 o C. The yield of transfectants in response to 10 pulses was up to 1000 per 106 cells (using 3.3 pg DNA per cell). The inßuence of the growth phase of the cells on the transfection yield and I or the subpopulation of the mouse L--ceU line used was shown. Furthennore the clone yield depended on the DNA per ceU ratio within a very small range. 1987 urn:nbn:de:bvb:20-opus-63497 Institut für Pharmakologie und Toxikologie OPUS4-6046 Wissenschaftlicher Artikel Zimmermann, U.; Stopper, Helga Elektrofusion und Elektropermeabilisierung von Zellen : Eine neuartige Methode der Biotechnologie zur gezieltenVeränderung der genetischen Eigenschaften von Zellen No abstract available 1987 urn:nbn:de:bvb:20-opus-63514 Institut für Pharmakologie und Toxikologie OPUS4-6032 Wissenschaftlicher Artikel Stopper, Helga; Zimmermann, U.; Neil, G. A. Increased efficiency of transfection of murine hybridoma cells with DNA by electropermeabilization 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. 1988 urn:nbn:de:bvb:20-opus-63488 Institut für Pharmakologie und Toxikologie OPUS4-6910 Wissenschaftlicher Artikel Stopper, Helga; Zimmermann, U.; Wecker, E. High yields of DNA-transfer into mouse L-cells by electropermeabilization no abstracts available 1985 urn:nbn:de:bvb:20-opus-82408 Institut für Pharmakologie und Toxikologie