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Five Ways to Produce siRNAs

Validated siRNAs: Silencer Validated siRNAs are single siRNA duplexes that have been verified experimentally to reduce the expression of individual human genes. Each siRNA has been shown to reduce target gene expression by at least 70% forty-eight hours post transfection by real time RT-PCR. Most of the siRNAs were found to reduce target gene expression by 90% or more. Every Silencer Validated siRNA strand is purified by HPLC and subjected to rigorous quality control measures. The result is the highest quality siRNA with a sequence verified to reduce gene expression.


Method #2: In Vitro Transcription
siRNAs can be readily prepared by in vitro transcription. siRNAs produced by this method are considerably less expensive than their chemically synthesized counterparts, making them a more cost-effective choice for screening siRNA sequences. In addition, they can be produced more quickly than chemically synthesized siRNAs. Once template deoxynucleotides are obtained, the procedure takes about 24 hours, with little hands on time. Disadvantages of this method include the limited scale up potential (although each reaction produces enough siRNA for hundreds of transfections) and the fact that it requires more hands-on time from the researcher compared to chemically synthesized siRNAs which can simply be purchased. It should be noted that in vitro transcribed siRNAs work as well as chemically synthesized siRNAs and usually at lower concentrations -- 0.520 nM vs. 50100 nM concentration per transfection (Figure 1).

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Screening siRNA sequences
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P8B1...
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