cotransfect or bicistronic?

Soenke Behrends behrends at plexus.uke.uni-hamburg.de
Fri Mar 10 15:20:20 EST 2000


Dear netters,

Is it fair to say that when you transfect pEGFP with green
fluorescence only and another vector with the same features
with your gene of interest that the fluorescence leads you
with some likelihood to the cells expressing you gene of
interest (e.g. for electrophysiology)?

i have heard (but do not have good scientific reference)
for the notion that when you transiently transfect cells 
(say with 10% of cells being transfected) you mostly 
get the same 10% of cells cotransfected with the another
similar vector. 

Fusion protein would of course a good alternative but fluorescence
is not strong enough in our case. 

Or is the use of a bicistronic vector (IRES marker control vector, 
Clontech) a good idea? If the ideas from the first paragraph
are true I would be happy to get around the cloning involved. 

Thanks for your time and help
Soenke






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