In transient transfection, the introduced nucleic acid exists in the cell only for a limited period of time and is not integrated into the genome. Therefore, the new gene will not be replicated. The effects of the foreign gene within the cell last only a short amount of time (usually several days), after which the foreign gene is lost through cell division or other factors.
Cells that are frequently used for transient transfection are HEK293 or CHO cells. Large volume transient transfection of HEK293 and CHO cells adapted to suspension culture has addressed the need to obtain amounts of recombinant protein without the time-consuming and labor-intensive creation of stable cell lines.
Whether you are focused on discovery, production of targets, or characterization of interesting gene products or antibodies, our one-stop transient transfection expression will help optimize the expression and purification of your target gene.
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First of all you need a cell line which is easy to transfect and will allow high product titers. Large volume transient transfection of HEK293 and CHO cells adapted to suspension culture has addressed the need to obtain high amounts of recombinant protein. Some transfection protocols require serum-free conditions for optimal performance, since serum can interfere with many commercially available transfection reagents.
DNA quality strongly influences the results of transfection experiments. The best results are achieved when plasmid DNA of the highest purity is used for transfection.
Cells should be transfected at 50–80% confluency. Too few cells will cause the culture to grow poorly. Too many cells results in contact inhibition.
Ensure that cells are active and at least 90% of them survive before transfection. The cell density is controlled at 1.5-2×106 cells/mL.
Several methods for transfection of foreign (target) gene have been developed using liposomes, calcium phosphate, electroporation, non-liposomal lipids, DEAE-dextran, microinjection and retroviral transfection.
The selection of transfection technology can strongly affect the transfection efficiency. In ideal conditions, transfection should be quick and easy to operate, providing high efficiency and repeatable results with minimal cytotoxicity.
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