Figure 5. Reduction of GAPDH Gene Expression in Normal Human Dermal Fibroblasts-Neonatal after Electroporation using siPORTer-96 Electroporation Chamber. siRNA targeting GAPDH or a non-targeting siRNA (1.0 g) were electroporated into Normal Human Dermal FibroblastsNeonatal (an adherent primary cell type) using the siPORTer-96 Electroporation Chamber (Ambion) powered by a Bio-Rad Gene Pulser Xcell pulse generator. Samples 148 (shown) were transfected with Silencer GAPDH siRNA (Ambion) while samples 4996 were transfected with Silencer Negative Control #1 (Ambion) (data not shown). 48 h post-transfection, cells were harvested and target mRNA levels were analyzed by real-time RT-PCR. 18S rRNA levels were used to normalize GAPDH expression. Remaining Gene Expression (%) was calculated as a percentage of gene expression compared with the averaged value of cells transfected with the negative control siRNA.
Many siRNA applications require delivery of
hundreds to thousands of siRNAs to cells with high efficiency
while minimizing toxicity. Chemical reverse transfection with
siPORT NeoFX Transfection Agent efficiently and
reproducibly delivered siRNAs to immortalized, adherent cells
in 96 and 384 well plates. In