http://www.scientistsolutions.com/t12788-luc_+_+gfp_stability.html Life Science Discussion en-us sci7feed@sci7.nojunkorherepleasespam.com hourly 1 2005-05-05T12:00+00:00 http://www.scientistsolutions.com/t12788-luc_+_+gfp_stability.html it will take some time, I have just started cloning. But I let you know my experience.
Andi
]]> Tue, 06 Oct 2009 01:47:56 GMT http://www.scientistsolutions.com/t12788-luc_+_+gfp_stability.html I don't think so.... :-( Unless the plasmid somehow got integrate during the DNA replication cycle...
But I may be wrong... If you have tried it out, I am very curious to know... :-)
]]> Mon, 05 Oct 2009 18:03:08 GMT http://www.scientistsolutions.com/t12788-luc_+_+gfp_stability.html yes, you are right. The usage of stable transfected cells would be the better option, but I have to test several constructs before I can make stable transfected cells.
Luckily, the cells I have to use for these experiments stop dividing before the tranfection and they are "healthy" for the next two weeks. Do you think that reporter gene vectors are stable for 2 weeks in these cells?
Andi
]]> Mon, 05 Oct 2009 09:40:05 GMT http://www.scientistsolutions.com/t12788-luc_+_+gfp_stability.html If you really have to measure the activity 2 weeks after transfection, I personally don't think it is a good idea to carry out the transfection in a transient manner. This is because there is very high chance that the untransfected cells will eventually catch up and dominate the cell population, thus you end up many cells without your plasmids and compromising your results.
Can you try to select for stable cell lines? Once you get the stable lines, you can always mak...]]> Mon, 05 Oct 2009 06:52:10 GMT http://www.scientistsolutions.com/t12788-luc_+_+gfp_stability.html I like to perform transient transfections with luciferase- and GFP-plasmids. Because of the time-protocol I have to meassure the activity of the reporter genes after 2 weeks. Has anyone an idea or experience with the stability of these vectors?
Thanx in advance,
Andi
]]> Mon, 05 Oct 2009 04:34:16 GMT