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Comprehensive Controls for Optimizing RNAi Experiments and Ensuring Transfection Efficiency

To ensure successful RNAi experiments, it's crucial to optimize transfection conditions and control for efficiency. The BLOCK-iT™ Transfection Control Kit and RNAi Basic Control Kit provide essential reagents. Early experiments should include unrelated or scrambled siRNA/shRNA controls and both positive and negative controls to assess cell health post-gene knockdown. Intermediate experiments should build on early controls, using multiple d-siRNA/siRNA/shRNA targeting the same gene. Full publication-quality controls include a dose-response curve and the optional analysis of global gene expression.

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Comprehensive Controls for Optimizing RNAi Experiments and Ensuring Transfection Efficiency

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  1. Controls for RNAi experiments PRIOR TO FIRST EXPERIMENTS and FOR ALL EXPERIMENTS optimize transfection conditions and control for transfection efficiency. The BLOCK-iT™ Transfection Control Kit and RNAi Basic Control Kit contain reagents to do this. • EARLY EXPERIMENTS: • Unrelated or scrambled siRNA/shRNA control • Positive and Negative control The BLOCK-iT™ RNAi Basic Control Kit can help assess cells health following gene knockdown • INTERMEDIATE EXPERIMENTS: • All controls in “EARLY EXPERIMENTS” • Multiple d-siRNA/siRNA/shRNA to same target gene • Show effect at mRNA level (LUX™primers/QRT-PCR or Northern) • “Stress response” gene monitoring (PKR or 5’OAS pathways) • OPTIONAL: Western analysis The BLOCK-iT™ RNAi Stress Response Kit can help monitor for activation of toxic pathways • FULL SET OF CONTROLS (publication quality): • All controls in “INTERMEDIATE EXPERIMENTS” • Dose-response curve • Rescue of phenotype • OPTIONAL: Global gene expression- arrays “Whither RNAi” Nature Cell Biology (2003) vol 5(6): 489-490

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