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High Throughput siRNA Electroporation

adherent cells like NHDF-neo cells.

Figure 3. GAPDH Gene Silencing and Cell Viability Under Various Electroporation Conditions. siRNA (1 g) targeting GAPDH (Silencer GAPDH siRNA, Ambion) or a Negative Control siRNA (Silencer Negative Control #1 siRNA, Ambion) were electroporated into normal human dermal fibroblast cells (an adherent primary cell type) using the siPORTer-96 Electroporation Chamber. Four samples were electroporated at a time, either with an siRNA targeting GAPDH (two samples) or with the negative control siRNA (two samples). The voltage was varied between 200 and 600 V for either 250 or 400 sec as labeled. 24 hours after electroporation, 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 samples transfected with the negative control siRNA (bars). Cell viability was measured with ViaCount Cell Viability assay (Guava) (diamonds).


Summary

When used with Bio-Rad's Gene Pulser Xcell pulse generator, Ambion's siPORTer-96 Electroporation Chamber enables efficient high throughput electroporation of siRNA. Coupled with the siPORT siRNA Electroporation Buffer, the electroporator provides highly efficient siRNA delivery into primary and other hard-to-transfect cell types with minimal cell mortality. Standard sample volume is 44 l, and enough siPORT siRNA Electroporation Buffer (12 X 1.5 ml) is supplied with the siPORTer-96 Elec
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