Has anybody good experience with stable co-transfection in 3T3 fibroblasts

Helfried Mallow helfried.mallow at vkl.uni-regensburg.de
Thu Jun 20 11:18:43 EST 1996


High,

I need someone who has good experience in stable transfection of 3T3 
fibroblasts. 

Problem : I have to transfect two constructs into 3T3-L1 preadipocytes 
(fibroblasts). One repressor construct of 6.7 kb and and expression construct 
of 7.8 or 8.8 kb. Transfection should be performed with Maxifectin, a liposome 
based method. 

Difficulty: I have not much time to find out detailed and optimal transfection 
conditions. I need an established method, in order to save time and get a 
satisfactory transfection efficiency.

Selection has to be performed with Hygromycin B and G418

Special questions : What are the optimal concentrations of those two 
antibiotics in a double selection? How long will the selection last? I am 
working in a serum-based system. 

I am an absolute beginner referring to transfection. 

My e-mail adress is : manfred.maier at vkl.uni-regensburg.de

If you or somebody else in your environment has got experience, please contact 
me.

Thanks a lot for your efforts

Manfred Maier 



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