3 replies to this topic

[ammahew]

Hi everyone,

Can anyone suggest a protocol for transfection of HT29 cells with siRNA using Lipofectamine 2000?

I have successfully transfected HCT116 cells, but have been told HT29s are more difficult to transfect. I'll try using the same protocol as the HCT116s initially, but I was wondering if anyone had any experience transfecting HT29s - are they more difficult to transfect? Will I need to increase the siRNA/Lipofectamine concentration, or transfect for longer?


Thanks!

[bob1]

You will need to optimize the transfection for your purposes; each cell line is different.  I would recommend however, that you get some lipofectamine RNAiMax instead of 2000, it is a lot less toxic and seems to work really well for siRNA transfection.

[ammahew]

Ok, I think I might actually have some RNAiMax somewhere from an old lab, thanks!

[vitalgene]

i would suggest a reverse trasfection for HT29 cells.