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stealth siRNA optimisation

stealth siRNA optimisation - Protocols and Methods Forum

stealth siRNA optimisation - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 07-11-2008, 09:27 AM
Scott Brown
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Default stealth siRNA optimisation



Hi,

I have been trying to use some invitrogen stealth siRNA to knock down the function of the glucocorticoid receptor and expression of BCDIN3.
I am using a nucelofector for transfecting the siRNA, it recommends using 2uM [siRNA], I have tried using 5um and 10uM as well and have achieved better results (almost 80% knockdown).

I always thought high concentraions of siRNA can be toxic to cells, can anyone advice on the mechanism of action responsible for this toxicity, is their any limit to how much siRNA i can expose my cell line to or is this a typical scenario when optimising siRNA knockdown.

Thanks

Scott Brown
PhD Candidate
Children's Cancer Institute of Australia

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optimisation , sirna , stealth


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