How to make a stable cell line using suspension cells - (Mar/21/2005 )

hi, everyone,
I am going to transfect suspension cells and make a stable cell lines. But I have no experience for this. Could any one teach me that?

thanks !

hi
for transfection of suspension cell line you may see this topics :

http://www.protocol-online.org/forums/inde...935&#entry10935

well i would say that culturing cells in the media qsupplemented with selection antibiotic would give you good results for establishing cell line.
Here is a protocol that may help you.

best time for adding G418

antibiotics during transfection

tell us if you want more precisions... There are also topics in cell biology forum taht may interest you...
fred

Hi,
Thank you so much for your help. It is really important for me. I will post my result in the near future.

Thank you for the protocol of selecting transfected cell. But there is a picture in it I can't acess. Is any way I can get it?

Thanks again.

hi

i can't get the picture too...

Directly not related to this thread but wanted to know whether GFP expression looks fainter in cells in suspension compared to attached cells? The infections which I do to attached cells look bright green but the cells in suspension are green but too faint!