Technik » Problem with GFP

Hi guy,

I stably transfected my cell with plasmid containing 3FLAG-my insert- IRES-GFP. The cells were first selected by puromycine and because I could not detect well the GFP sign under microscope, I did the FACS to sort the GFP positive cells. After FACS sorting the GFP positive cells, I grow my cells on medium containing puromycine, but when I rechecked my cells again 1 week after by the FACS machine, almost all the cells showed negative GFP.

What is the problem and how I can improve to get more GFP positive cells?


Please help....


I didn't find the right solution from the Internet.
References
http://molecularbiology.forums.biotechni...hp?f=17&t=31878
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