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
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,
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.
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
Source:Page: All 1 2 3 Related biology technology :1
. Efficient DNA transfection of primary CNS neurons using TransMessenger
. TransMessenger Transfection Reagent Handbook3
. TransMessenger Transfection Reagent4
. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN
siRNA and TransMessenger Transfection Reagent*5
. Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell
. Optimizing Transfection Conditions for Studying Signal Transduction Pathways7
. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells8
. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal
. Versatile Transfection Reagent Offers Low Toxicity and Consistent
. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells11
. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection
of Mammalian Cells