( only) The ratios...)Transfection Reagent Selector Kit Handbook

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The ratios of DNA to Enhancer provided in the selector protocol should not be changed. The ratio of DNA to Enhancer is 1 g of DNA to 8 l of Enhancer. Efficient condensation of DNA with Enhancer is only determined by the mass quantity of DNA. (Cell line or plasmid size do not influence the DNA-to-Enhancer ratio).

Ratio of Transfection Reagent to DNA

The ratio of Transfection Reagent (l) to DNA (g) is an important factor to optimize for every new cell line and DNA construct used. Optimal binding of Transfection ReagentDNA complexes to the negatively charged groups (e.g. sialylated glycoproteins) on the cell surface requires a slightly net positive charge. For Effectene Reagent, the overall charge of the Transfection ReagentDNA complex is determined by the ratio of Transfection Reagent to DNAEnhancer mixture. For SuperFect Reagent, the overall charge of the Transfection ReagentDNA complex is determined by the ratio of Transfection Reagent to DNA. Figure 2 represents a pipetting scheme for optimizing the ratio of DNA to Transfection Reagent for transfection of adherent cells or suspension cells in 6-well plates.

Incubation period with Transfection ReagentDNA complexes
For SuperFect Reagent:

For adherent cells, the length of incubation of transfection complexes with cells should be optimized by varying the incubation time within a range of 116 h. Optimal results are typically obtained by choosing periods of 23 h.

For suspension cells, experiments have shown that in most cases removal of SuperFectDNA transfection complexes is not necessary. However, if cytotoxic
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