TransMessenger Transfection Reagent

For efficient mRNA transfection

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

TransMessenger Transfection Reagent (0.5 ml)

Cat. No. / ID: 301525

For 60 transfections in 6-well plates or 80 transfections in 12-well plates

The TransMessenger 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

Highly efficient transfection
Reproducible results ensured by stringent quality control
Efficient transfection of primary neuronal cells

Product Details

TransMessenger Transfection Reagent is a ready-to-use, lipid-based reagent for RNA transfection of eukaryotic cells. Transfection of cells with RNA rather than DNA offers new possibilities for transfection experiments. Cell lines successfully transfected with TransMessenger Reagent include 293T, Jurkat, and Vero. This product can also be used for RNA transfection of primary human fibroblasts, rat cardiomyocytes, and rat neuronal cells.

Performance

TransMessenger Reagent provides a fast and easy procedure and high transfection efficiencies (see figure " TransMessenger Reagent with HeLa cells"). Since the amount of RNA is a critical factor for successful transfection, we recommend optimizing the amounts of RNA and TransMessenger Transfection Reagent for every cell type–RNA combination (see figure " Amount of RNA and TransMessenger Reagent vs. transfection efficiency"). To facilitate this, the reagent is provided with guidelines for optimization together with starting points for optimization in different cell-culture formats.
TransMessenger Transfection Reagent is highly suited for use in functional cardiac cell studies using neonatal rat ventricular cardiac cells.
Cardiomyocytes from cryopreserved, dissociated neonatal rat ventricular cardiac cells (R-CM-561 [QBMCellScience.com]) display excellent viability, pharmacology, morphology, and connectivity, as well as contractile and electrical activity necessary for use in functional screening (see video Transfection control: Functional Cardiomyocytes Beating). To evaluate the suitability of using TransMessenger Transfection Reagent in functional cardiac cell studies, cells were transfected with a GFP-expression vector 4 hours after plating, and were fixed 5 days post-transfection. Use of TransMessenger Transfection Reagent resulted in an excellent, 60% survival rate post-transfection with a transfection efficiency of 10–13%. All cardiomyocytes displayed beating post-transfection (sees videos GPF-Positive Cardiomyocytes Beating Post-Transfection and Functional Cardiomyocytes Beating Post-Transfection), indicating functional cells. The number of transfected cardiomyocyte versus non-cardiomyocyte cells was evaluated and results demonstrate that a high number of cardiomyocyte and non-cardiomyocyte cells were transfected. Equivalent morphologies between transfected and non-transfected cells were also observed (see figure Cardiac cell studies).

See figures

TransMessenger Reagent with HeLa cells.

Amount of RNA and TransMessenger Reagent vs. transfection efficiency.

Cardiac cell studies.

Principle

TransMessenger Transfection Reagent is the first reagent specifically developed for transfection of cells with RNA. The reagent is a lipid-based formulation that is used in conjunction with a specific RNA-condensing enhancer and an optimized buffer. RNA molecules are condensed by the enhancer and then coated by TransMessenger Reagent for efficient transfer into eukaryotic cells. Strict quality control is performed to test for absence of RNase activity, lot-to-lot consistency, and low endotoxin levels (≤10 EU/ml). Our rigorous standards eliminate reagent variables that can adversely affect the efficiency of RNA transfection.

Procedure

All TransMessenger Reagent components are provided as ready-to-use solutions. To generate TransMessenger–RNA transfection complexes, simply mix your RNA with Enhancer R and Buffer EC-R and incubate for 5 minutes at room temperature, then add TransMessenger Reagent and incubate for a further 5–10 minutes. The complexes are mixed with medium and added directly to the cells. Following a 3 hours incubation, the medium is changed and the cells are incubated until they are ready for analysis.
Optimal transfection results are achieved using high-purity RNA that is free of DNA, proteins, and other contaminants. RNA purified with RNeasy is highly recommended.

Applications

Transfection of cells with RNA rather than DNA offers new possibilities for transfection experiments. Transfected RNA sequences are expressed in the absence of transcription, and in a promoter-independent manner. In addition, protein expression usually occurs sooner following transfection of RNA rather than DNA.

RNA transfection with TransMessenger Transfection Reagent can be used for:

Studies of cells not efficiently transfected with plasmid
DNA Direct studies of RNA function

Supporting data and figures

Amount of RNA and TransMessenger Reagent vs. transfection efficiency.

Optimization experiments were performed in CHO-K1 cells. Cells (2 x 10⁴⁾ were seeded in quadruplicate into 96-well plates and transfected 24 hours later with an in vitro-transcribed CAT-encoding RNA using [A] increasing amounts of RNA with 1.5 µl TransMessenger Reagent and [B] increasing amounts of TransMessenger Reagent with 0.25 µg RNA, as described in the TransMessenger Transfection Reagent Handbook. CAT activity was measured 24 hours post-transfection.

Specifications

Features	Specifications
Applications	Direct studies of RNA function
Features	Transfection with RNA. Efficient transfection of neuronal cells.
Controls	Not included
Nucleic acid	RNA
Cell type	Eukaryotic cells
Number of possible transfections	80 transfections in 12-well plates / 0,5 ml reagent
Technology	Lipid-based formulation in conjunction with a RNA-condensing enhancer
Transfection type	Transient transfection, co-transfection

Resources

Search for transfection data by nucleic acid, cell line, and transfection reagent. Our database contains data from researchers like yourself who have shared their experimental results with us.

Transfection protocols for specific cell types and plate formats that save you the time and effort of adapting existing protocols to fit your requirements. Simply select the cell type, nucleic acid, and culture format to receive a QIAGEN transfection protocol to print out or download in convenient PDF format.

For transfection of eukaryotic cells with RNA and siRNA

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

Download Safety Data Sheets for QIAGEN product components.

The following procedure is for co-transfection of adherent cells using siRNA and plasmid DNA in one well of a 24-well plate. This procedure is provided as a starting point for optimization of siRNA and plasmid DNA co-transfection in mammalian cells using TransMessenger™ Transfection Reagent.

FAQ

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

Yes, please follow the Supplementary Protocol 'Guidelines for co-transfection of adherent cells with siRNA and plasmid DNA using TransMessenger Transfection Reagent' (TFP21).

In addition, we have also the following Supplementary Protocol for the Attractene Transfection Reagent 'Fast-Forward cotransfection of adherent cells with siRNA and DNA in 24-well plates using Attractene Transfection Reagent'.

FAQ ID -969

Unfortunately, we do not have any data for the transfection of siRNA into insect cells. However, we know about customers who have successfully used Effectene Transfection Reagent for the transfection of plasmid DNA into insect cell lines S2 and Sf9 (see FAQ 397). Since Effectene Transfection Reagent and HiPerFect Transfection Reagent for the transfection of eukaryotic cells with siRNA are both lipid based, there is a chance that HiPerFect will work with insect cells. It is especially suitable for transfection of very low siRNA concentrations, reducing the risk of cell toxicity and off-target knockdown effects.

Alternatively, you can try RNAifect or TransMessenger Transfection Reagent, also lipid-based, for the transfection of siRNA into S2 cells.

We would appreciate any feedback on your results in case you want to give it a try. Please visit our Transfection Cell Survey page to submit your feedback.

FAQ ID -1046

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

No, we do not recommend to use RNAiFect for the transfection of plasmid DNA. The RNAiFect Kit does not contain Enhancer which is necessary to form condensed complexes before transfection of DNA into the cell. TransMessenger Transfection Reagent would be the right choice as it allows co-transfection of siRNA and plasmid DNA. Access the protocol via the following link: Co-transfection of adherent cells with siRNA and plasmid DNA using TransMessenger Transfection Reagent

Alternatively, you can use the Attractene Transfection Reagent with the QIAGEN Protocol "Fast -Forward cotransfection of adhernet cells with siRNA and DNA in 24-well plates using Attractene Transfection Reagent".

FAQ ID -515