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Choose the Right Transfection Agent for Your RNAi Experiment

Figure 4. Delivery of siRNA by siPORT Lipid and siPORT Amine Induces RNAi. The indicated cell types were transfected with GAPDH siRNA or negative control siRNA using either siPORT Amine or siPORT Lipid Transfection Agent. RNA was isolated with the RNAqueous Kit and gene silencing effects were monitored by Northern blot using the NorthernMax-Gly Kit. Bars indicate gene expression levels 48 hours after delivery of the gene-specific siRNA relative to negative control wells.


Optimizing siRNA Transfection Parameters
By choosing the right transfection agent and optimizing cell density, transfection agent amount, and siRNA concentration, high levels of transfection efficiency can be achieved in many cell types. For developing an optimal siRNA transfection protocol in your experimental system, Ambion has combined the siPORT Amine and siPORT Lipid agents with a GAPDH- targeting siRNA and a negative control siRNA in the Silencer Transfection Kit. After transfection of the positive control, reduction in GAPDH activity can be monitored by immunofluorescence or Western blot with Ambion's monoclonal antibody to GAPDH, or by Northern blot, RT-PCR or RPA. Alternatively, siRNA uptake can be monitored directly by fluorescently labeling the si
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