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Five Ways to Produce siRNAs

s address delivery of both RNA-based siRNAs and DNA-based expression vectors by transfection and electroporation and provide tips for optimizing this step. Poor delivery efficiency is one of the main reasons RNAi experiments fail (or give false negative results) and deserves much attention.

Analysis of Gene Silencing
Detection and quantitation of gene silencing can be assayed by measuring mRNA or protein levels, or both. The articles on pages 18 and 21 discuss some of these methods. Controls that will help determine siRNA specificity are described in Designing Controls for siRNA Experiments.


Summing It Up
The use of RNAi to induce gene silencing is an exciting new method for creating loss of function phenotypes. Table 1 on page 5 summarizes the five different methods for producing siRNAs. More information about each of these methods can be found throughout this issue. Additional details, including protocols, can be found on the RNAi Resource page at www.ambion.com/RNAi.


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Ordering Information
Cat# Product Name Size 1620 Silencer siRNA Construction Kit 15 siRNA
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Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

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3. Cenix-Designed siRNAs for 95% of Human and Mouse Genomes
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
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8. Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success
9. Deliver siRNAs Into Primary Cells
10. Optimize Transfection of siRNAs for RNAi
11. RNAi as a Tool for Mammalian Gene Analysis: Applications of siRNAs
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