Can any one share their knowledge about the best way of transfecting of K562 with GFP plasmid please?

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Noor Shamkhi

thank you so much

Kamal Shaik Fakiruddin

Dear Shamkhi,

We have few researchers in our lab trying to transfect siRNA-GFP into K562 cells using common lipid-based methods (e.g Lipofectamine, Fugene, Turbofect) but the efficiency was very low. The efficiency goes up only when we switch the electroporation-based method (e.g. Nucleofection, Neon). I think lipid-based transfection works well on adherent cells but not for floating (non-adherent) cells as most of the lipid complex would just sink to the bottom of the flask leaving most cells happily floating on top.

Hopes that may help,. good luck.. ☺

Dear Fakiruddin,

thank you for your answer, I have to try the options

regards

I will be grateful, if anyone share their knowledge about a good method for fixing and permeabilizing cells for inserting dyes inside the nucleus and then use the flow cytometry to measuring.