I've done transient knockdown with siRNA using Hiperfect and I wondered- should I have used the same concentration of non silencing control as of the siRNA for the target gene. As it happens there was a 5 fold difference in concentration. Don't ask me why! I just noticed it. .
I fully agree with the points of Venkatesh. I use Lipofectamine 2000, and it works pretty well with our conditions.
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The more controls- the better: untreated cells, positive control, negative control (for some important experiments and siRNA screening with many targets we actually use several neg controls of completely different seq), all at the same concentration. I actually like (when possible) to run experiments with three siRNA concentrations- to see the dose effects (eg 0.1, 1, 10 nM in cells)
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