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TransMessenger Transfection Reagent

For transfection of cells with siRNA in RNAi procedures Features and benefits
  • Efficient transfection of siRNA for effective gene silencing
  • Quick and easy setup
  • Ready-to-use reagent
  • Tested for absence of RNase activity
Principle TransMessenger Transfection Reagent facilitates effective gene silencing in RNAi procedures by providing efficient transfection of siRNA into eukaryotic cells using a fast and easy procedure (see figure "Efficient Transfection of Primary Neuronal Cells Using QIAGEN siRNA and TransMessenger Reagent"). The reagent is a lipid-based formulation that is used in conjunction with a specific RNA-condensing enhancer and an optimized buffer to form siRNA-TransMessenger Reagent complexes that are efficiently transferred 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 siRNA transfection. Efficient Transfection of Primary Neuronal Cells Using QIAGEN siRNA and TransMessenger Reagent Primary neuronal cells from rats were transfected with a control nonspecific siRNA or an siRNA directed against microtubule-associated protein 2 (MAP2) using TransMessenger Transfection Reagent. Two days after transfection, cells were fixed, permeabilized, and stained using mouse monoclonal anti-MAP2 antibodies and Alexa Fluor 488-conjugated goa t anti-mouse secondary antibody (a-MAP2) and Alexa Fluor 594-conjugated phalloidin, which binds to f-actin and serves here as an expression control.

Data kindly provided by A.M. Krichevsky and K.S. Kosik, Brigham and Womens Hospital, Harvard Medical School, Boston, MA (from Krichevsky, A.M., and Kosik, K.S. (2002) RNAi functions in cultured mammalian neurons. Proc. Natl. Acad. Sci. USA 99, 11926. Copyright 2002 National Academy of Sciences, U.S.A.)

Procedure All TransMessenger Reagent components are provided as ready-to-use solutions. To generate TransMessenger-RNA transfection complexes, simply mix your siRNA with Enhancer R and Buffer EC-R and incubate for 5 minutes at room temperature, then add TransMessenger Reagent and incubate for a further 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 siRNA. The QIAGEN siRNA Oligo Synthesis Service offers custom siRNA oligos, a range of library siRNAs that target specific genes, non-silencing control siRNAs, and a set of siRNAs targeting genes linked with cancer; the Cancer siRNA Oligo Set Version 1.0.
Since the amount of siRNA used is a critical factor for successful transfection, we recommend optimizing the amounts of siRNA and TransMessenger Transfection Reagent for every cell type-siRNA combination. To facilitate this, the reagent is provided with guidelines and starting points for optimization of RNAi experiments.

App lications TransMessenger Reagent has been used to mediate successful RNAi in several studies using different cell types. For an up-to-date list of references and the latest developments in QIAGEN products for RNAi visit the Transfection Tools site at www.qiagen.com/transfectiontools.


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