Transfecting from all types of nucleic acids with a very high efficiency

TransFect Plus Transfection-Reagent: Manual


Transfect Plus reagent is our latest generation of transfection reagent. It allows transfecting all types of nucleic acids with a very high efficiency. Due to its formulation, Transfect Plus reagent delivers a large quantity of nucleic acids leading to higher protein expression compared to other transfection reagents. It is fully biodegradable and does not interfere with cellular mechanisms.  Consequently, high cell viability is maintained in every experiments and any potential secondary effect is avoided

Transfect Plus reagentcan be used to realize any transfection applications: co-transfection, reverse transfection, high-throughput screening…
 
• Superior protein expression level
• Great transfection efficiency
• Biodegradable - Avoid secondary effects
• For all nucleic acids types
• Serum compatible
• Simple, rapid and easy-to-use

Outstanding transfection reagent

Transfect Plus reagentoutperforms all other transfection reagents in efficiency and transgene expression level. In addition, an important cell viability is maintained even for sensitive cells.

High DNA transfection efficiency

GFP expression in different cell lines



Percentage of transfected cells




Exceptional protein expression in hard-to-transfect cells
B-galactosidase expression



Superior cells transfection efficiency

Outperforms all tested transfection reagents


High cell viability

Effects of various transfection reagents on cytotoxicity




General Considerations

The instructions given below represent sample protocols that were applied successfully with a variety of cell lines. Optimal conditions may vary depending on the nucleic acid, cell types, size of cell culture dishes and presence or absence of serum. Therefore, the amounts and ratio of the individual components (DNA and TransfectPlus  ) may have to be adjusted to achieve best results since each cell line has a particular optimal transfection reagent / nucleic acids ratio. As a result, we suggest you to optimize the various transfection parameters as described in section 3.7) Optimization Protocol. The following recommendations can be used as guidelines to quickly achieve very good transfection and high transgene expression level. As a starting point, we recommend to use 4 µL of TransfectPlus / 1 µg of DNA. TransfectPlus can be used in the presence or in the absence of serum. You can use your routine culture medium for the transfection, except during preparation of the TransfectPlus / DNA complexes (see 3.3 below).

• Cells should be healthy and assay during their exponential growing phase. The presence of contaminants (mycoplasma, fungi) will considerably affect the transfection efficiency. The cell proliferating rate is a critical parameter and the optimal confluency has to be adjusted according to the cells used. We recommend using regularly passaged cells for transfection and avoid employing cells that have been cultured for too long (> 2 months). Generally, siRNA transfection requires lower cell density than DNA transfection.

• Nucleic acids should be as pure as possible. Endotoxins levels must be very low since they interfere with transfection efficiencies. Moreover, we suggest avoiding long incubation time of the DNA/RNA solution in buffers or serum free medium before the addition of TransfectPlus reagent to circumvent any degradation or surface adsorption.

• Antibiotics. The exclusion of antibiotics from the media during transfection has been reported to enhance gene expression levels. We did not observe a significant effect of the presence or absence of antibiotics with the TransfectPlus reagent and this effect is cell type dependent and usually small.

• Materials. We recommend using polypropylene tubes to prepare the DNA and transfection reagent solutions but glass or polystyrene tubes can also be used.

A protocol used for other transfection reagents should never be employed for TransfectPlus and inversely. Each transfection reagent has its own molecular structure, biophysical properties and concentration, which have an important influence on their biological activity.
 

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