( any lab can take advantage ...)Perform RNAi Library Screens on Any Budget

Figure 1. Overview of RNAi Experiments Using the Silencer CellReady siRNA Library System. Step 1A: Search the Ambion website and choose a Silencer CellReady siRNA Library. Step 1B: Optimize siRNA transfection conditions for your cell line using the Silencer CellReady siRNA Transfection Optimization Kit. Step 2: Perform reverse transfection experiments in triplicate using the Silencer CellReady siRNA Library from Step 1A and the optimized conditions determined in Step 1B. Step 3: Assay cellular phenotype (e.g., cellular proliferation or apoptosis). Compare results from samples transfected with each gene-specific siRNA to those transfected with negative control siRNA. Step 4: Validate hits detected in Step 3 by a secondary assay.

Data Above: Gene Expression Results After Apoptosis and Cell Proliferation Screening of Silencer CellReady siRNA Plates Specific for CMGC Kinases. The data in Step 3 are from an actual screening experiment where CellReady sets of siRNAs, representing three individual siRNAs per target and targeting the CMGC subset of human kinases, were used to transfect HeLa Cells in triplicate using siPORT NeoFX. A confirmed hit was defined as any target gene that had two or more single siRNAs that induced a change in signal by > two standard deviations in the same direction as compared to cells transfected with Silencer Negative Control #1 siRNA. Signals were normalized by cell number. (A) Cells were assayed with alamarBlue (Biosource) 72 hr post transfection to identify kinases, that when silenced, affected cell pro
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