siRNA transfection reagent - (Dec/08/2006 )

hi
I use to have aplsmide with my shRNA sequenc in it.
I want to try to oder synthetize siRNA and transfect my celll with it.

what type of transfectant should I use for a doublestrand RNAoligo?

any advice?

jm

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We commonly use Lipofectamine2000 and in special cases Oligofectamine.

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i would contact some companies that make siRNA transfection reagents (promega, ambion, dharmacon etc.) and ask for a sample packet so you can optimise your transfection relative to the reagent as these things can be quite expensive!

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well a colleague routinely use lipofectamine and Plus reagent.

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lipofectamine 2000

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it's f...ing toxic to my cells... anyone knows something about Jetsci transfection reagent?
thanks
Toni

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try lower the amount of lipofectamine that u r using for transfection. may b this could help.

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no way, I tried DNA/siRNA:lipofectamine2000 ratios ranging from 1:1 to 1:5..., with the lower ratios the result was very low transfection effiency, with the higher ratios good efficiency but also toxicity
toniX

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what quantity of lipo/siRNA are you using?
I read that 125nM of siRNA is routinely used...

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50nM of siRNA and 2,5 ul of lipo in a 12 well...
, I used this concentration because I have observed the maximum inhibition in my system...

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