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TransMessenger Transfection Reagent (0.5 ml)
Cat. No. / ID: 301525
For 60 transfections in 6-well plates or 80 transfections in 12-well plates
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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 verses 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).
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 verses 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
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 hour 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 and Oligotex mRNA Kits is highly recommended.
Optimal transfection results are achieved using high-purity RNA that is free of DNA, proteins, and other contaminants. RNA purified with RNeasy and Oligotex mRNA Kits 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 | |
| Features | |
| Controls | |
| Nucleic acid | |
| Cell type | |
| Number of possible transfections | |
| Technology | |
| Transfection type |
Resources
Transfection Protocols (2)
Kit Handbooks (1)
Brochures & Guides (3)
Quick-Start Protocols (1)
Supplementary Protocols (1)
Safety Data Sheets (1)
FAQ
Do you have transfection data for QIAGEN Transfection Reagents?
Do you have a protocol for co-transfection of adherent cells with siRNA and plasmid DNA?
How can I transfect siRNA into insect cells such as Drosophila melanogaster S2?
What is the recipe for 1x PBS solution?
Can I use RNAiFect for co-transfection of plasmid DNA and siRNA?