stealth siRNA optimisation

Scott Brown via methods%40net.bio.net (by SBrown from ccia.unsw.edu.au)
Fri Jul 11 04:27:40 EST 2008


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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