( ...)Optimizing electroporation parameters

Other important parameters for optimizing electroporation.

• Field strength
• Pulse length
• Number of field pulses
• Adjustment of the electroporation buffer
• Influence of DNA/RNA
• Influence of temperature
• Influence of the cell density

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Influence of the DNA/RNA The transfection efficiency of electroporation can be affected by the concentration, the purity and the size of the molecules used.

a) Influence of nucleic acid concentration:
With the optimal electroporation parameters (osmolarity, voltage, pulse length), the quality of the results obtained with plasmid concentrations between 5 g/ml and 20 g/ml is usually satisfactory. The efficiency of the transfection may be raised by increasing the nucleic acid concentration, but only within a limited concentration range. Tests with various different cell lines have shown that only in very few cases (e.g. when large plasmids were used) do plasmid concentrations in excess of 20 g/ml lead to an increase in the transfection rate (see Fig. 1.).

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