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621 results sorted by
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The quality and the concentration of DNA used for Nucleofection™ in general plays a central role for the efficiency of gene transfer. We strongly recommend using endotoxin free prepared DNA. Endotoxin free Kits are available from several suppliers....
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The duration time of a program is dependent on the number of the program. The duration is also dependent on how long the Nucleofector™ Device has been turned on as well as the number of Nucleofections you have done in the last couple of minutes. When...
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Insect cells are cultivated at lower temperatures than E.coli or mammalian cells (S2 cells at 24°C, Sf9 cells at 28°C). Therefore they need a dedicated incubator with appropriate temperature.
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Vectors with a CMV promoter [e.g. pmaxCloning™, Lonza, Cat. No. VDC-1040] typically work fine for transient protein expression in mammalian cells, like HEK293, CHO or NSO. For insect cells, like Sf9 or Sf21, vectors with insect promoters are...
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The recommended amount of DNA per 100 µl reaction is 0.5µg-5 µg. No carrier is needed.
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The optimal incubation time ranges from 2-days to 6 days depending on various factors like the stability of the protein in the medium, potential toxicity of the protein, cell density and stability of the plasmid in the nucleus. To check the timepoint...
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No. The electrical parameters provided by the Nucleofector™ Device are optimized for the cuvettes contained in the Nucleofector™ Kits. The cuvettes are single-use only. Using the cuvettes more than once will result in higher cell death and lower...
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For many cell lines in suspension, especially sCHO, conventional transfection methods are very limited in efficiency. For protein production, suspension cells are favored over adherent cells because they are much easier to cultivate. Unlike...
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No, it seems that the beads bound to the cell surface do not have any influence on Nucleofection™. Larger magnetic beads (i.e. > 1µm) may disturb the Nucleofection™ process and negatively influence cell viability and efficiency.
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The generation of a stable clone often requires six months or more due to selection procedures and adaption to serum-free conditions after transfection. With the help of the Nucleofector technology suspension cells can be transfected directly in a...
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