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Silencer Validated siRNAs

dated siRNAs were effective when transfected at concentrations below 100 nM, though the data point to the fact that poor transfection efficiency can be overcome by using higher concentrations of siRNA (Figure 3).

Figure 2. Effectiveness of Silencer Validated siRNAs at Various Concentrations. The indicated siRNAs were mixed with siPORT Lipid (Ambion) and the resulting complexes were added to HeLa cells in 24 well plates at the final concentration of siRNA shown. Forty-eight hours after transfection, RNA from the treated cells was recovered using the RNAqueous-MAG Total RNA Isolation Kit (Ambion) and reverse transcribed using the RETROscript Kit (Ambion). Target cDNA levels were measured by real-time PCR using SYBR Green assays. The expression of the target genes in the transfected cells was compared to cells transfected with an equal concentration of the Silencer Negative Control #1. Input cDNA in the different samples was normalized using real-time data for 18S rRNA. The bar graphs represent an average of three data points.

Figure 3. siRNA Activity in Various Cell Types. HeLa, HepG2, MCF7, and SK-N-AS cells in 24 well plates were transfected with the indicated siRNAs at various concentrations. Forty-eight hours after transfection, RNA from the samples was isolated using the RNAqueous-MAG Total RNA Isolation Kit (Ambion) and target gene expression was measured by real-time RT-PCR using SYBR green assays. The expression of the target genes in the transfected cells was compar
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