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X-tremeGENE siRNA Transfection Reagent

Polymer reagent for delivering siRNA to common cell lines

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Synonym(s):
siRNA, transfection reagent

grade

for molecular biology

Quality Level

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])

manufacturer/tradename

Roche

concentration

1 mg/mL

technique(s)

transfection: suitable

shipped in

wet ice

storage temp.

2-8°C

General description

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.

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.

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.

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).

Other Notes

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

Legal Information

X-tremeGENE is a trademark of Roche

Storage Class Code

12 - Non Combustible Liquids

WGK

nwg

Flash Point(F)

does not flash

Flash Point(C)

does not flash


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J S Ungerstedt et al.
Proceedings of the National Academy of Sciences of the United States of America, 102(3), 673-678 (2005-01-08)
This study examines the basis of resistance and sensitivity of normal and transformed cells to histone deacetylase inhibitor (HDACi)-induced cell death, specifically the role of caspases and thioredoxin (Trx). An important attribute of HDACis is that they induce cancer cell
Overexpression of microRNA-1 impairs cardiac contractile function by damaging sarcomere assembly.
Ai J
Cardiovascular Research, 95(3), 385-393 (2012)
Yves Decrem et al.
International journal for parasitology, 38(5), 549-560 (2007-10-26)
Two cDNAs coding homologous putative metalloproteases (Metis 1 and Metis 2, expected molecular weights of 55.6 and 56.0kDa, respectively) were identified from the hard tick Ixodes ricinus. The expression of Metis genes was induced in salivary glands during tick blood
Hak-Ryul Kim et al.
Lung cancer (Amsterdam, Netherlands), 60(1), 47-56 (2007-11-17)
Heme oxygenase-1 (HO-1) is highly expressed in various tumor tissues and plays an important role in tumor cell growth through anti-oxidative and anti-apoptotic effects. Herein, we demonstrate that A549 cells express high levels of HO-1, Nrf2, and NF-kappaB compared to
Mei Zhan et al.
Experimental hematology, 35(7), 1015-1025 (2007-06-26)
MicroRNAs (miRNAs) are an abundant class of small noncoding RNAs that regulate diverse cellular functions by sequence-specific inhibition of gene expression. We determined miRNA expression profile during erythroid differentiation and putative roles in erythroid differentiation. The expression profile of 295

Articles

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 mammalian cells without having to resort to classical gene knockout techniques.

Transfection is the introduction of DNA, RNA, or proteins into eukaryotic cells and is used in research to study and modulate gene expression. Thus, transfection techniques and protocols serve as an analytical tool that facilitates the characterization of genetic functions, protein synthesis, cell growth and development.

This brief webinar provides an overview of what transfection is and the methods that are used to introduce DNA or RNA into eukaryotic cells.

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 enabled expression and isolation of recombinant proteins in large scale.

Protocols

X-tremeGENE™ siRNA Transfection Reagent Protocol & Troubleshooting

Protocols for Transfecting Common Cell Lines with X-tremeGENE™ Transfection Reagents

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