METAFECTENE® SI
The siRNA and miRNA transfection reagent for eukaryotic cells
Highlights
- Specific optimization of lipid composition for RNA transfections (siCOM Technology)
- Integrated RMA and TOP Technology
- Efficient knockdown and low siRNA volume
- Rapid protocol: two interdependent assays within one week*
- Protocol extremely suitable for screening processes
METAFECTENE® SI is a lipid compound designed and optimized especially for siRNA and miRNA transfection (siCOM Technology). The characteristic second-generation features of Biontex's patented and proven RMA and TOP technologies deliver high knockdown rates, low RNA concentration and very low toxicity, enabling them to be used for all cell lines and primary cells.
Learn more about siCOM Technology
Sequence of expression of GFP in HeLa-GFP cells transfected with METAFECTENE® SI and siRNA directed against the mRNA of the GFP. Clearly visible is the reduction in GFP expression to a maximum knockdown at 72 hours. Note that GFP has a long half-life, resulting in this delay in knockdown.
siCOM-Technology
(siRNA adjusted Composition)
Unlike conventional transfection reagents for DNA transfection, reagents for the transfection of siRNA and miRNA do not need to transport the DNA into the cell nucleus. The development of METAFECTENE® SI was thus primarily aimed at ensuring the maximum endocytosis volume in the lipoplex plus rapid and complete release of RNA in the cytosol.
A broadly based screening process enabled a highly active lipid composition precisely aligned to siRNA and miRNA transfection to be identified from a comprehensive library of lipids. In this context, a specifically optimized buffer composition for lipoplex formation– the SI buffer – and a significantly simplified protocol were developed. These individual features combine to form a unique composition which enables outstanding siRNA and miRNA transfection rates to be achieved.
This new development also permits a reverse protocol which is simplified and one day shorter than conventional protocols. In addition, the product uses a predefined ratio of RNA to lipids, rendering complex optimization of the lipoplex compound unnecessary. Because of the time saving from four days to three, two successive result-dependent transfections can be performed within a single working week*.
Screening processes benefit both from this timesaving and from the fact that the lipoplex formation, and thus all pipetting steps, take place in the well of a multiwell culture plate, eliminating the need for rinsing or transfer. siRNA or miRNA transfection can thus be performed during screening with the maximum efficiency and ease.
Learn more about RNA Interference
RNA Interference
RNA Interference (RNAi) is a natural regulatory mechanism in gene expression, controlled by short-chain RNA with approx. 21 – 28 nucleotides which are complementary to the mRNA of a target protein and can be found in all eukaryotic cells.
A distinction is made here between siRNA and miRNA, which is formed in the case of siRNA from doublestrand RNA (e.g. endogenic from transposons or exogenic from viruses) and, in the case of miRNA, from hairpin-structure endogenic RNA (from own pri-miRNA genes) by means of enzymatic cleavage with an endoribonuclease (Dicer).
These small RNA strands are bound in the cytosol by a ribonucleoprotein complex – the RISC complex (RNA induced silencing complex), the RNA double strand is separated and the leading strand is presented. When a complementary mRNA strand binds to the leading strand, it is broken down or deactivated by the RISC complex, thus preventing expression of the target protein and down-regulating the corresponding gene (knockdown).
The discovery of this mechanism, known as "gene silencing", has been a major research milestone in recent years, and is a powerful tool in the field of proteomics and research into functional relationships between genes.
| Product Specifications |
|
| Application |
Transfection of eukariotic cells with siRNA or miRNA |
| Formulation |
Cationic lipids with colipids in water |
Content
| Lipid formulation / sufficient quantity of SI buffer
|
| Assays |
Approx. 1000 (96-well) or approx. 250 (24-well) per 1 ml reagent |
| Shipping |
At room temperature |
| Storage |
≤ –15°C |
| Shelf Life |
1 year (after date of delivery) |
* At max. 48-hour incubation period between lipoplex addition and evaluation.
Prices
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