Double Transfection problems? - (Oct/17/2005 )
Hello...I've been having problems with double transfections and am getting a little frustrated.
I have a small peptide coupled with CFP (~250 aa) and another gene with YFP thats about 1500 aa. When I try to do a double transfection I only see one or the other with fluorescence (I have appropriate dichroics).
Even more bizarre I have made a polyclonal cell line under selection pressure with the YFP vector. I try to transiently transfect the CFP and still the problem. 95% of the ones expressing YFP have no CFP. 95% of the cells expressing CFP have no YFP. There is a very small population that has both vectors at very very very small expression levels.
Any thoughts?
-DavidK-
QUOTE (DavidK @ Oct 17 2005, 03:01 AM)
Hello...I've been having problems with double transfections and am getting a little frustrated.
I have a small peptide coupled with CFP (~250 aa) and another gene with YFP thats about 1500 aa. When I try to do a double transfection I only see one or the other with fluorescence (I have appropriate dichroics).
Even more bizarre I have made a polyclonal cell line under selection pressure with the YFP vector. I try to transiently transfect the CFP and still the problem. 95% of the ones expressing YFP have no CFP. 95% of the cells expressing CFP have no YFP. There is a very small population that has both vectors at very very very small expression levels.
Any thoughts?
I have a small peptide coupled with CFP (~250 aa) and another gene with YFP thats about 1500 aa. When I try to do a double transfection I only see one or the other with fluorescence (I have appropriate dichroics).
Even more bizarre I have made a polyclonal cell line under selection pressure with the YFP vector. I try to transiently transfect the CFP and still the problem. 95% of the ones expressing YFP have no CFP. 95% of the cells expressing CFP have no YFP. There is a very small population that has both vectors at very very very small expression levels.
Any thoughts?
what abouth the promoter competition?
-Jusu-