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High Throughput siRNA Electroporation

ontrol #1 siRNA, Cat# 4611). Data are shown for the GAPDH siRNA-treated samples. Normal expression levels of GAPDH were reduced by a mean of 94.8% with a coefficient of variance of 18.9%. Variability is attributed to steps in the procedure such as pipetting error, measurement error, and assay error, as well as true biological variability.

Figure 2. Reduction of GAPDH Gene Expression in NHDF-neo Cells. siRNA (1 g) was electroporated into NHDF-neo (normal numan dermal fibroblast cells, an adherent primary cell type) using the siPORTer-96 Electroporation Chamber. Samples 1-48 were transfected with an siRNA targeting GAPDH (Silencer GAPDH siRNA, this figure), and samples 49-96 were transfected with a negative control siRNA (Silencer Negative Control #1 siRNA, data not shown). 48 hours after transfection, the cells were harvested and analyzed by real-time RT-PCR for GAPDH expression levels. 18S rRNA levels were used to normalize GAPDH expression. Remaining Gene Expression was calculated as a percentage of GAPDH gene expression compared with the averaged value from cells transfected with the negative control siRNA.


Figure 3 shows results of experiments used to identify optimal electroporation conditions for NHDF-neo primary cells. An siRNA targeting GAPDH and a Negative Control siRNA were again used for this experiment. Both remaining GAPDH expression levels and cell viability were measured. These data illustrate the impressive gene silencing that can be achieved by delivering a potent siRNA with the siPORTer-96 Electroporation Chamber while maintaining cell viability. A wide range of parameters are effective in delivering siRNA to
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