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X-tremeGENE siRNA Transfection Reagent

Polymer reagent for delivering siRNA to common cell lines

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Properties

Related Categories Cell Biology, Cloning and Expression, Life Science Research, Molecular Biology, Synthetic Biology,
grade   for molecular biology
usage   sufficient for 2,000 transfections (04476115001)
  sufficient for 400 transfections (04476093001)
packaging   pkg of 1 mL (04476093001 [1 mg/ml])
  pkg of 5 × 1 mL (04476115001 [5 x 1 mg/ml])
mfr. no.   Roche
shipped in   wet ice
storage temp.   2-8°C

Description

Features and Benefits

• Knock down gene expression over 90% in many different cell types.
• Maximize experimental flexibility with a single reagent for siRNA- and cotransfection-based gene-knockdown experiments.
• Produce meaningful results using a reagent that exhibits low cytotoxicity, ensuring that the cellular effects you observe are due to the transfected siRNA rather than the transfection procedure.
• Work with or without serum, avoiding medium changes (e.g., to serum-free medium) before or after transfection.;

X-tremeGENE siRNA Transfection Reagent is a proprietary blend of lipids and other components, free of animal products.

General description

Download the CofA, SDS and IFU specific to your product lot and format

Short interfering RNA (siRNA) can be directly introduced into cells by transfection. A lipid-based transfection reagent, such as X-tremeGENE siRNA Transfection reagent, can provide a convenient, reliable, and efficient vehicle for delivering siRNAs into animal cells, enabling the study of cellular and functional consequences of gene knockdown.
This innovative reagent forms a complex with siRNA, as well as with mixtures of siRNA and plasmid DNA (cotransfection), and efficiently delivers the nucleic acids into animal cells to induce gene silencing. Transfection is achieved in just a few steps: mix and incubate diluted transfection reagent with diluted siRNA, then add this complex to cells. Because X-tremeGENE siRNA Transfection Reagent functions exceptionally well in the presence or absence of serum and demonstrates low cytotoxicity, it can be used without media changes. The product is animal-component free.

Contents

Solution filtered through 0.2 μm pore size membrane, supplied in polypropylene tubes.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Quality

Activity assay: X-tremeGENE siRNA Transfection Reagent (1 - 2.5 μl) is combined with siRNA (0.1 - 0.35 μg) that is specific for the HPRT housekeeping gene. The mixture is used to transfect HEK-293 cells (in a monolayer, 30 - 50% confluent) in the presence of 10% fetal bovine serum (FBS). Following transfection, the decrease of HPRT mRNA cells is determined with the LightCycler® Real-Time PCR System. A knockdown efficiency of 70 - 95% is typically observed when mRNA is measured.

Cytotoxicity analysis: HEK-293 cells are exposed to siRNA/X-tremeGENE siRNA Reagent complexes for 72 hours in the presence of serum, without a media change. Cytotoxicity is then tested by analyzing the cells with the WST-1 Cell Proliferation Reagent (Roche).

Application

X-tremeGENE siRNA Transfection Reagent efficiently delivers short interfering RNA (siRNA) into many commonly used cell types including HeLa, NIH 3T3, HEK-293, CHO-K1, and COS-7, and several hard-to-transfect cell lines, such as HT29, a human adenocarcinoma cell line.

Legal Information

X-tremeGENE is a trademark of Roche

Price and Availability


Milli-Q® Water Purification Solutions
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis

Protocols & Articles

Articles

A Versatile new Lipofection Reagent for co-transfection of DNA and RNA: X-tremeGENE™ siRNA Transfection Reagent

Small inhibitory RNAs (siRNAs) have become the focus of interest in many laboratories. For the first time, these molecules offer an easy way to knock down the expression of selected genes in mammalia...
Keywords: Antibiotics, Diagnostic, Gene expression, Genetics, Transfection, Western blot

Molecular Biology Handbook

Molecular Biology is the science that aims to understand biological activity at the molecular level. These biological activities usually involve the plant or animal cell, and the nucleic acids and pr...
Keywords: Cell culture, Cell signaling, Cloning, Detection methods, Electrophoresis, Gel electrophoresis, Gene expression, Genetics, Molecular biology, Molecular biology techniques, Nucleic acid hybridization, Nucleic acid purification, Polymerase chain reaction - quantitative, Protein electrophoresis, Purification, Transfection, Western blot, transformation

Protein Expression Systems

The development of genetic engineering and cloning has opened many possibilities of expression and isolation of heterologous proteins for research purposes. Considerable advances in technology have e...
Keywords: Amplification, Antibiotics, Cell disruption, Cloning, Culture media, Gene expression, Genetic, Genetics, Glycosylations, Molecular biology, Purification, Pyrogens, Transfection, transformation

Roche Transfection Database

Roche X-tremeGENE™ Transfection Reagents have been used to transfect hundreds of different cell lines, primary cells & stem cells. To find the right product for your cells, use the Selection Guide be...
Keywords: Apoptosis, Cancer, Clinical, Cloning, DNA replication, Gene expression, Immunoprecipitation, In Situ hybridization, Individual protein Immunoprecipitation, Ion-exchange chromatography, Methylations, Microscopy, Nucleic acid hybridization, Polymerase chain reaction, Recombination, Transcription, Transfection, Western blot

Transfection Best Practices

The work of scientific research is an art which requires good attention to detail.  The quality of transfection is highest when these commonly used best practices and notes on optimization are employ...
Keywords: Antibiotics, Apoptosis, Growth factors, Molecular biology, Purification, Transfection

X-tremeGENE™ General Recommendations

Note: The above data was produced using either a GFP-encoding pcDNA3.1 plasmid or a luciferase-encoding pCI plasmid, both with the cytomegalovirus (CMV) promoter. These recommendations are guidelines...
Keywords: Cancer, Cellular processes, Diagnostic, Gene expression, Gene knockdown, Transfection

X-tremeGENE™ siRNA Transfection Reagent: A Powerful Tool for Antisense Inhibition of miRNA in Human Cells with miRCURY Knockdown Probes

MicroRNAs (miRNAs) are an endogenous class of RNA molecules comprising 19-25 nucleotides that regulate the suitability or transflational efficiency of mRNAs. The mature miRNAs are processed from long...
Keywords: Apoptosis, Cancer, Cell proliferation, Diagnostic, Gene expression, In Situ hybridization, Nucleic acid hybridization, Transfection

Protocols

Protocols for Transfecting the Most Common Cell Lines with X-tremeGENE™ Transfection Reagent

Plate CHO-K1 cells at a density of 1.5 X 104 cells/well. Plate cells in a volume of 100 μl complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (65 – 75% confluency). ...
Keywords: Diagnostic, Gene expression, Transfection

X-tremeGENE™ siRNA Transfection Reagent Protocol & Troubleshooting

siRNA IN PRIMARY LUNG FIBROBLASTS TRANSFECTION Roche has no data for this application, but there is a report from a customer, briefly described below. Wild rat lung fibroblasts were successfully cotr...
Keywords: Gene expression, Growth factors, Microarray Analysis, Transfection, Western blot

Peer-Reviewed Papers
15

References

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Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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