siRNA in non-adherent cell lines? - Tips please (Mar/03/2006 )

I'm going to try knockdown in a non-adherent cell line, I've had success with oligofectamine in adherent cell lines but wanted to ask if anyone had managed using oligofectamine for transfecting non-adherent cell lines? Or can anyone recommend a better method?

-Sarah F-

hi
i use lipofectamine foer adherent cells and for non adherent ones. Protocol is the same regarding time of tranfection and ratio DNA/lipocomplex. I use twice DNA quantity per same amount of cells.

-fred_33-

Cheers, I'll try that one out

-Sarah F-

some threads may be useful too...
In search you type suspension cells. I've listed some..

QUOTE (kevin77 @ Aug 17 2005, 04:49 PM)

I have worked using siRNA with Jurkat cells and K562 cells in my previous lab. For such suspension cells, normally lipid-based transfection reagents have presented very low transfection effiencies. I have tested OpenBiosystem Arrest-In and Roche x-tremeQ2 (for K562, Jurkat and HeLa), both got <10% transfection efficiency, and Arrest-In is more toxic.
For that reason, we turned to eletroporation-Amaxa Nucleofector, after optimization, for early passage Jurkat cells (<10), we could get >90% efficiency.
Good Luck!
kevin

http://www.protocol-online.org/forums/inde...wtopic=9210&hl=
http://www.protocol-online.org/forums/inde...wtopic=5711&hl=
http://www.protocol-online.org/forums/inde...wtopic=1969&hl=

-fred_33-

That's a great help I'll look into trying out some of those methods.

-Sarah F-