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Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success

titated by real-time PCR.
5) Data are normalized using 18S rRNA levels.
6) The extent of target gene knockdown is expressed as a percent of mRNA remaining in cells treated with the gene-specific siRNA
compared to cells treated with a negative control siRNA (Silencer Negative Control siRNA #1).

Figure 2. Silencer siRNA Validation Data Generated Using Applied Biosystems TaqMan Gene Expression Assays. The indicated Silencer Validated siRNAs were transfected into HeLa Cells at 30 nM. RNA was isolated 48 hours later and analyzed by one-step qRT-PCR using the appropriate TaqMan Gene Expression Assay (results were normalized for input RNA amount using real-time data for 18S rRNA). The inset graphs show the reduction in target gene expression compared to cells transfected with an equal concentration of Silencer Negative Control #1.


Optimizing siRNA Transfection
Before siRNA experiments can be conducted to study gene function, analyze biological pathways, or validate drug targets, optimized siRNA delivery conditions must be identified and/or verified. The best way to test, optimize, and validate siRNA transfection or electroporation conditions is to use a highly effective, verified siRNA to an appropriate target and to monitor decreases in mRNA levels induced by delivery of the siRNA. Ambion uses qRT-PCR as the method of choice, and Applied Biosystems TaqMan Gene Expression Assays provide validated, easy-to-use primer-probe sets for this purpose (F
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Related biology technology :

1. Silencer Validated siRNAs
2. Cenix-Designed siRNAs for 95% of Human and Mouse Genomes
3. Five Ways to Produce siRNAs
4. Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection
5. IMPROVE SILENCING SUCCESS: siRNAs THAT WORK BETTER!
6. Optimizing Chemical Transfection and Electroporation of siRNAs
7. Delivering siRNAs to Difficult Cell Types
8. Deliver siRNAs Into Primary Cells
9. Optimize Transfection of siRNAs for RNAi
10. RNAi as a Tool for Mammalian Gene Analysis: Applications of siRNAs
11. Test More siRNAs for Less
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