Transfection with Effectene - (Feb/07/2005 )

Hi,

I encounter a problem in transfecting HEK 293 A cell. The first time I transfect with Qiagen Effectene reagent and the cell is pretty good and transfection of GFP shows that it's successful. However, I did transfection again with the same reagent, but right after I transfect the cell, they float. And It does not seem to be blown up.

Plus, I am always using antibiotics and serum containing medium cause the protocol siad"can contain". And previously, it works.

Thanks a lot

I presume you do treat the plates with glycine or poly-lysine. THis sticks them down to the plate. 0.1mg/ml treamtents for a few hours then wash off with sterile water 3x is good.

hi
for transfection, you can use serum during transfection. But 293cells do not need serum for achieving good efficiency.

But may i advice you to not use antibiotics during transfection. your cells are enough damaged by the mixture lipid molecule DNA, and do not need to deal with antibiotics too....