MISSION ® siRNA

Validation and Guarantee

 
 
How do you validate your Predesigned siRNAs?
The answer for this is two-fold. All of our MISSION Predesigned siRNAs are first validated computationally, using the Rosetta Algorithm.
 
We offer a subset of our siRNA as Validated MISSION Predesigned siRNA. The Validated MISSION Predesigned siRNAs are validated functionally as outlined below.
 
Transfection reagents: Various transfection reagents were used to functionally validate our siRNA constructs.
siRNA concentrations: Various concentrations have been used, however, the most common is 50 nM and this would comfortably cover over 60% of the validation work we've completed. The 50 nM was agreed upon primarily because that was the concentration Rosetta used when first validating their algorithm. Subsequently, we have looked at various concentrations of our siRNA and confirmed performance at much lower concentrations.
Cell lines examined: Nearly all of our validation testing has been completed in the HeLa cell line.
Formats: 96-well format was used.
Replication: We always conducted minimally three biological replicates and three technical replicates for each construct examined. Additionally, viability testing was done on nearly all validation experiments.
Times to harvest mRNA: Most measurements were obtained at 24 hours post-transfection.
Methods to assess knockdown: Branched DNA technology or qRT-PCR.
 
Do you guarantee that your siRNAs will knock down my gene?
Sigma-Aldrich guarantees that two out of three siRNA duplexes per target gene will achieve knockdown efficiencies of greater than or equal to 75% for our MISSION Predesigned siRNAs. Custom designed siRNAs are not guaranteed. Please see our full guarantee.
 
How should I choose the 3 siRNA so that I have the benefit of your guarantee?
Sigma-Aldrich guarantees that two out of three siRNA duplexes per target gene will achieve knockdown efficiencies of greater than or equal to 75% for our MISSION Predesigned siRNAs. Custom designed siRNAs are not guaranteed. When getting started on a new experiment, we recommend choosing the top three Rosetta Algorithm designs.