Can I use the Nucleofector Technology for RNAi applications? How do I start?
Yes. The Nucleofector Technology can be easily applied for any RNAi substrate (siRNA, shRNA, miRNA). You can use the same conditions described in the cell type specific Optimized Protocol for DNA vectors (cDNA, shRNA or miRNA expressing plasmids) or oligonucleotides (siRNA, miRNA inhibtors). Prior to beginning comprehensive experiments multiple parameters associated with experimental design need to be optimized: Selection of appropriate controls (non-targeting siRNA, siRNA targeting a house-keeping gene, untreated cells). Determination of minimum effective siRNA amount for your target gene in your cell type of interest. Determination of the optimal analysis time point (kinetics of down-regulation depend on target gene, cell type and analysis level). Identification of a suitable and robust analysis method (i.e. mRNA, protein or phenotypic analysis). Please click on the associated link for example data and recommendations.