Can I transfect HEK293 cells using siRNA purchased form Dharmacon and use Lipofectamine reagent instead of DharmaFECT? I have been using electroporation for the transfection, but it seems like HEK cells do not tolerate this method, so I am trying to find something less "toxic" and more toleratable.
Thank you for any advise!
Transfection agents should work independently of the siRNA (or equivalent short oligos) as long as you follow the protocol. Based on personal experience, all polymeric/lipid based reagents including the most advanced lipofectamine or the old diethylaminoethyl dextran will have some cytotoxicity. Electroporation is superior if used in the correct conditions. Having said that, adherent cells will not like to be electroporated in suspension, there are plate electrodes which can be used for adherent cells electroporation and they work perfectly fine without the need to detach the cells.
Hasan Issa Thank you for the help. I will try to use Lipofectamine for the transfection of HEK cells. I transfect a leukemic cell line using the electroporation under the same condition as HEK293, and it doesnt have any severe effect on cell viability.
Hello!
HEK cells are very easy to transfect and you definitively do not need to electroporate them. You can use any reagent for siRNA on the market. lipid-based or polymer, they should all work.
We for example use these cells to test/validate each lot of our Viromer reagents, it's routine work.
I would simply suggest to think also about the price and the amount of siRNA you may need. Lipofectamine/s are quite expensive...
Best, Sandra
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