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Choose the Right Transfection Agent for Your RNAi Experiment

he presence or absence of serum. siPORT XP-1 is simple to use -- reagent/DNA complexes can be added directly to growing cells in serum-containing culture media, and the media can be left unchanged until the time of assay.

An example of the plasmid transfection efficiencies that can be obtained with siPORT XP-1 is shown in Figure 1. In this figure, a plasmid designed to express -galactosidase was transfected into HeLa cells using siPORT XP-1 and the cells were analyzed by microscopy. Transfection rates of 70-80% have been achieved with HeLa cells as shown by the blue staining.

Figure 1. siPORT XP-1 Efficiently Delivers Plasmid DNA. HeLa cells were transfected with a plasmid expressing -galactosidase using siPORT XP-1. 48 hours after transfection, the cells were treated with X-gal and analyzed by microscopy. The blue color indicates -galactosidase expression.


In gene silencing experiments, high transfection efficiencies are essential in order to observe the down regulation of genes by Northern or Western analysis, or any other technique that analyzes the entire cell population, containing both transfected and nontransfected cells. In Figure 2, siPORT XP-1 was used to transfect COS-7, HeLa, and DU145 cells with a plasmid, pSilencer 2.0-U6, engineered to express an siRNA to GAPDH. Silencing effects were analyzed at the mRNA level by Northern blot 72 hours after transfection. In all three cell lines, GAPDH mRNA levels were reduced >80%, demonstrating the effectiveness of plasmid transfections by siPORT XP-1 in these three cell lines.

Figure 2. Gene Silencing in Mult
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