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Choose the Right Transfection Agent for Your RNAi Experiment

iple Cell Lines with an siRNA Expression Plasmid Transfected with siPORT XP-1. pSilencer 2.0-U6 was engineered to express an siRNA to GAPDH and then transfected into COS-7, HeLa, and DU145 cells with siPORT XP-1. 72 hours after transfection, RNA was isolated with the RNAqueous Kit and GAPDH mRNA levels were analyzed by Northern blot using Ambion's NorthernMax-Gly Kit. NC=no construct, ie cells transfected with transfection agent alone.


Transfecting PCR products, such as those prepared using Ambion's Silencer Express siRNA Expression Cassette Kit, can be challenging as many plasmid transfection agents are not suitable for transfection of these shorter, linear dsDNAs. However, XP-1 is also efficient at introducing SECs and other PCR products into a range of mammalian cell types. Protocols for transfecting SECs are distinct from those for transfecting plasmid expression vectors and are outlined in a separate section of the siPORT XP-1 Protocol. Figure 3 shows the use of siPORT XP-1 for transfecting SECs targeting GAPDH into five different cell lines. GAPDH mRNA levels were reduced by >90% in three of the five cell lines, and >75% in the remaining two.

Figure 3. SECs Delivered with siPORT XP-1 Induce Gene Silencing. siRNA Expression Cassettes (SECs) were prepared with the <
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