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Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success

"validated," to reduce target mRNA levels by a predetermined amount. Functional testing of a large number of siRNAs designed using a particular siRNA design algorithm is also necessary to prove that the algorithm used to design the siRNA accurately predicts effective siRNAs.

Ambion, and partner Cenix BioScience, chose qRT-PCR to test Ambion's Silencer Validated siRNAs and to systematically validate more than 1100 siRNAs to verify the effectiveness of Cenix's siRNA design algorithm (this algorithm was used to design all of Ambion's Silencer Validated and Pre-designed siRNAs; see siRNA Design: It's All in the Algorithm). Figure 1 illustrates the workflow used to validate siRNAs. Using this method, siRNAs that are determined to reduce their target mRNA level by 70% or more are considered "validated" and are subsequently made available as Silencer Validated siRNAs. Figure 2 shows a small subset of validation data generated at Ambion using TaqMan Gene Expression Assays to quantitate target mRNA levels.

Figure 1. Validation of Ambion's Silencer Validated siRNAs. The following procedure is used by both Cenix and Ambion to validate siRNAs:

1) Cells are plated in 96 well plates and grown for 24 hours.
2) Gene specific and negative control siRNAs are independently transfected in triplicate.
3) 48 hours later, RNA is extracted.
4) Target mRNA levels are quan
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Related biology technology :

1. Silencer Validated siRNAs
2. Cenix-Designed siRNAs for 95% of Human and Mouse Genomes
3. Five Ways to Produce siRNAs
4. Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection
5. IMPROVE SILENCING SUCCESS: siRNAs THAT WORK BETTER!
6. Optimizing Chemical Transfection and Electroporation of siRNAs
7. Delivering siRNAs to Difficult Cell Types
8. Deliver siRNAs Into Primary Cells
9. Optimize Transfection of siRNAs for RNAi
10. RNAi as a Tool for Mammalian Gene Analysis: Applications of siRNAs
11. Test More siRNAs for Less
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