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Controls for RNAi experiments
PRIOR TO FIRST EXPERIMENTS and FOR ALL
EXPERIMENTS optimize transfection conditions
and control for transfection efficiency.
EARLY EXPERIMENTS:
1.
Unrelated or scrambled siRNA/shRNA control
2.
Positive and Negative control
The BLOCK-iT™ Transfection Control
Kit and RNAi Basic Control Kit
contain reagents to do this.
The BLOCK-iT™ RNAi Basic Control Kit
can help assess cells health following
gene knockdown
INTERMEDIATE EXPERIMENTS:
1.
All controls in “EARLY EXPERIMENTS”
2.
Multiple d-siRNA/siRNA/shRNA to same target gene
3.
Show effect at mRNA level (LUX™primers/QRT-PCR or
Northern)
4.
“Stress response” gene monitoring (PKR or 5’OAS pathways)
5.
OPTIONAL: Western analysis
The BLOCK-iT™ RNAi
Stress Response Kit
can help monitor for
activation of toxic
pathways
FULL SET OF CONTROLS (publication quality):
1.
All controls in “INTERMEDIATE EXPERIMENTS”
2.
Dose-response curve
3.
Rescue of phenotype
4.
OPTIONAL: Global gene expression- arrays
“Whither RNAi” Nature Cell Biology (2003) vol 5(6): 489-490
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