Effectene Transfection Reagent

For DNA transfection of primary cells and sensitive cell lines

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Effectene Transfection Reagent (1 ml)

Cat. No. / ID: 301425

1 ml Effectene Reagent, Enhancer, Buffer; for 40 transfections in 60 mm dishes or 160 transfections in 12-well plates

Quantity

1 ml

4 x 1 ml

The Effectene Transfection Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

High efficiency in the presence of serum
Efficient transfection with low DNA amounts
Far lower cytotoxicity and gentler than many alternatives
Suitable for high-throughput screening

Product Details

Effectene Transfection Reagent is a nonliposomal lipid reagent for DNA transfection into a broad range of cell types. Due to its low cytotoxicity, Effectene Transfection Reagent is highly suitable for transfection of primary cells and many sensitive cell lines.

Performance

Higher transfection efficiencies of plasmid DNA are achieved with Effectene Transfection Reagent than with other reagents when used with the recommended procedures (see figure " High transfection efficiencies using Effectene Reagent"). Effectene Transfection Reagent is suitable for transfecting sensitive cell lines with oligonucleotides (see figure " Transfection of oligonucleotides using Effectene Reagent") and is particularly effective for primary cells (see figure " 40% transfection efficiency in primary cells"). Many cell lines and primary cells have been successfully transfected using Effectene Reagent; cell type-specific transfection protocols are available. Cytotoxicity is minimal because transfection with Effectene Reagent can be performed in the presence of serum and requires low amounts of DNA (see figure " Serum and DNA quantity vs. transfection efficiency").

The application of recombinant DNA technology to fields such as drug discovery and development has led to an increased need for high-throughput transfection. Transfection using Effectene Transfection Reagent requires low amounts of DNA and minimal handling. In addition, removal of transfection complexes is not required, making this reagent highly suitable for high-throughput screening.

See figures

High transfection efficiencies using Effectene Reagent.

Transfection of oligonucleotides using Effectene Reagent.

40% transfection efficiency in primary cells.

Serum and DNA quantity vs. transfection efficiency.

Principle

Effectene Transfection Reagent is an innovative non-liposomal lipid formulation that is used in conjunction with a special DNA-condensing enhancer and optimized buffer to achieve high transfection efficiencies. The enhancer first condenses the DNA molecules and Effectene Reagent subsequently coats them with cationic lipids providing a particularly efficient way of transferring DNA into eukaryotic cells. This feature ensures excellent reproducibility of transfection complex formation.

Procedure

The Effectene procedure has two steps. DNA is first mixed with Enhancer and a buffer that provides optimal salt conditions for efficient DNA condensation. This step requires just 2–5 minutes. Effectene Reagent is then added and the mixture is incubated for 5–10 minutes to allow Effectene–DNA complexes to form. The complexes are mixed with growth medium (which can contain serum and antibiotics), and added directly to the cells. The cells are then incubated until harvested and analyzed for gene expression (see flowchart " Effectene transfection procedure").

See figures

Effectene transfection procedure.

Applications

Effectene Transfection Reagent is suitable for transient and stable transfection of a broad range of cell types.

Supporting data and figures

High transfection efficiencies using Effectene Reagent.

Transfection efficiencies using Effectene Reagent and two commonly used lipid-based reagents were compared. Murine teratocarcinoma F9 cells (5 x 10⁵) were transfected in 6-well plates with a luciferase-reporter plasmid using optimized conditions based on the manufacturer's instructions for each reagent. Transfection efficiencies were determined by measuring luciferase activity 48 hours post-transfection, and are given as relative light units (RLU). (Data kindly provided by I. Clavereau, D. Petitprez, and I. Van Seuningen, Unité INSERM 377, Place de Verdun, Lille Cedex, France.)

Specifications

Features	Specifications
Applications	Plasmid transfection, protein overexpression, reporter studies
Technology	Non-liposomal lipid formulation in conjonction with a DNA-condensing enhancer
Number of possible transfections	160 transfections in 12-well plates / 1 ml reagent
Cell type	Eukaryotic cells (primary cells and sensitive cells)
Features	Non-liposomal lipid formulation, minimal cytotoxicity
Transfection type	Transient and stable transfection
Controls	Not included
Nucleic acid	DNA

Resources

The following protocol is optimized for transient transfection of CHO cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.

The following protocol is optimized for transient transfection of 293 cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.

The following protocol is optimized for transient transfection of COS-7 cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.

The next generation in lipid technology

Brochure detailing reagents for efficient and robust DNA and RNA transfection.

FAQ

The recommended amount of DNA being transfected should be split between the different plasmids. For example, if you normally transfect 5 µg of one plasmid, use 2.5 µg each for two plasmids (other proportions of the two plasmids can be used, as long as 5 µg total is maintained) as a starting point for optimization.

FAQ ID -124

QIAGEN Transfection Reagents have been used successfully with many different cell types. For your convenience, we have organized data from researchers who have shared their experimental results with us online in our Transfection Cell Database. Simply type your cell line of interest into the 'Search' field on this page, and find transfection results achieved with various QIAGEN Transfection Reagents. Please note that QIAGEN cannot verify data supplied from outside sources.

You can also submit your own transfection data and obtain a gift as appreciation. In addition you can find on our TransFect Protocol Database transfection protocols for specific cell types and plate formats.

FAQ ID -158

Customers have successfully transfected plasmid DNA into insect cell lines S2 and Sf9 using Effectene Transfection Reagent. For a reference for Drosophila S2 cells see QIAGEN News article Issue No. 4, 1999: 'Effectene Reagent yields high transfection efficiencies with Drosophila melanogaster S2 cells'.

Customer data for transfection of DNA into SF9 Spodoptera frugiperda cells using Effectene and SuperFect Transfection Reagent can be found in our Transfection Cell Database.

FAQ ID -397

The composition of 1x PBS solution is:

137 mM NaCl
2.7 mM KCl
4.3 mM Na₂HPO₄
1.47 mM KH₂PO₄

Adjust to a final pH of 7.4.

This solution is not supplied in any QIAGEN Kit, but is used in protocols for various QIAGEN transfection kits.

FAQ ID -1030

Effectene Reagent is a unique non-liposomal lipid formulation. Effectene Reagent is used in conjunction with an Enhancer and a DNA-condensation buffer (Buffer EC) to achieve high transfection efficiencies. In the first step of Effectene–DNA complex formation, the DNA is condensed by interaction with the Enhancer in a defined buffer system. Effectene Reagent is then added to the condensed DNA to produce Effectene–DNA complexes. The Effectene–DNA complexes are mixed with medium and directly added to the cells.

Effectene Reagent spontaneously forms micelle structures that show no size or batch variation, as found with preformulated liposome reagents. This unique feature ensures excellent reproducibility of transfection-complex formation. The process of condensing DNA molecules and then coating them with Effectene Reagent is a particularly effective way to transfer DNA into eukaryotic cells.

Broad cell line spectrum

Effectene Transfection Reagent has been used for transfection of a variety of different cell lines and primary cells, and yields significantly better transfection results than many widely used liposome-based transfection reagents. A searchable list of cell lines and primary cells successfully transfected using Effectene Reagent, as well as customer-developed protocols, is available at the Transfection Tools web site.

FAQ ID -184

The following paramenters can be optimized to increase transfection efficiency when using Effectene Transfection Reagent:

Optimize the Effectene Reagent to DNA ratio

If the ratio of Effectene Reagent to DNA is suboptimal, the overall charge of the complexes may be negative, neutral or strongly positive, which can lead to inefficient adsorption to the cell surface. Optimize the Effectene Reagent to DNA ratio according to the section " Transfection Optimization" of the Effectene Transfection Reagent Handbook.

Increase the amount of Effectene-DNA complex

If the transfection efficiency is lower than expected and cytotoxicity acceptably low, increase the overall amount of Effectene-DNA complexes. See the pipetting scheme in the section "Transfection Optimization" of the Effectene Transfection Reagent Handbook for details.

Optimize the incubation time for gene expression

Different cell types achieve maximal expression levels at different times post-transfection. This should be kept in mind when determining length of incubation after transfection. If the time point of maximal expression is not known for a particular cell line, a time course experiment may be necessary.

Consider vector influences

Factors such as the promoter, origin of replication, and plasmid size influence gene expression rate. The optimal quantity of plasmid DNA used for transfection is dependent on the expression rate of the plasmid.

Optimize cell density at the time of Effectene-DNA complex addition

If cell density is too high at the time of transfection-complex addition, cells may not be at the optimal phase of growth. This can lead to insufficient uptake of the complexes into the cells or insufficient expression of the gene of interest. For adherent cells, the optimal confluency at the time of transfection complex addition is normally 40-80%.

Use high-quality DNA Plasmid DNA used for transfection should be of high quality

Impurities present in the DNA preparation can potentially lower transfection efficiency. DNA should be purified using HiSpeed, QIAfilter, or QIAGEN Plasmid Kits or an equivalent method. For endotoxin-sensitive cell lines and primary cells, we recommend using DNA purified with EndoFree Plasmid Kits to ensure the highest transfection efficiencies.

FAQ ID -181