( electroporation conditions...)Deliver siRNAs Into Primary Cells

electroporation conditions for any cell type. For instance, successful delivery of siRNA into cells by electroporation depends on several critical parameters: voltage, pulse length, number of pulses, and siRNA concentration. The Cy3-labeled control siRNA makes it simple to monitor the uptake of siRNA into the target cells by fluorescence microscopy. The siPORT siRNA Electroporation Kit provides the means to dramatically simplify and speed up the electroporation optimization process, because a well-characterized siRNA targeting GAPDH and a widely used negative control siRNA and detailed optimization instructions are also included.

A Complete System for Optimization of siRNA Delivery

As seen in Figure 1, electroporation was used to deliver Cy3-labeled siRNA into human primary mesenchymal stem cells (hMSC) and rat neuronal pheochromocytoma (PC-12) cells. Almost every cell (>90%) contained detectable amounts of Cy3-labeled siRNA 24 hours after electroporation. This is a useful preliminary experiment to quickly screen multiple transfection conditions when working with new cell lines. A Cy3-labeled negative control siRNA comes with the kit for this purpose.

Figure 1. Electroporation of Primary Cells and Hard-to-transfect Neuronal Cells. A Cy3-labeled GAPDH siRNA (1.5 g) was added to primary Human Mesenchymal Stem Cells (hMSC) or rat neuronal pheochromocytoma (PC-12) cells in siPORT Electroporation Buffer (75 l) and electroporated using hMSC-specific or PC-12-specific parameters. Cells were fixed 24 hours afte
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