( and fungi should be avoided si...)Transfection Reagent Selector Kit Handbook

and fungi should be avoided, since it can drastically alter transfection results. Antibiotics can be included in the medium used for transfection with SuperFect Reagent and Effectene Reagent, and during subsequent incubation for gene expression.

Effect of serum

In contrast to many liposome transfection reagents, SuperFect Reagent and Effectene Reagent enable transfection in the presence of serum without lowering transfection efficiencies. Therefore, serum can be included in medium when incubating cells with transfection complexes and during subsequent incubation for gene expression. Use the same percentage of serum that cells have been adapted to in culture.

Vector construct

The type of transfection vector (plasmid DNA, RNA, PCR products, oligonucleotides) influences the transfection results. The configuration and size of the construct also determine the efficiency of transfection. Transient transfection is most efficient with supercoiled plasmid DNA. In stable transfection, linear DNA results in lower DNA uptake by the cells, relative to supercoiled DNA, but yields optimal integration of DNA into the host genome.

DNA quality

The best results are achieved when plasmid DNA of the highest purity is used for transfection. DNA purified with QIAGEN and QIAfilter Plasmid Kits is ideally suited for transfection of most cell lines. For transfection of endotoxin-sensitive cells, we recommend using DNA purified with EndoFree Plasmid Kits. These kits efficiently remove bacterial lipopolysaccharide molecules during the plasmid purification procedure, ensuring optimal transfection
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