2 replies to this topic

[Grenouille]

Hi,

I am planning to do an experiment in which I want to overexpress my protein of interest linked by T2A with GFP as marker. So my protein of interest and the gfp should get cleaved posttranslational.
Right now I am thinking about the best control for such an experiment. I think an empty vector (only with GFP) is better than mock but one could argue that in my construct is much more (longer) mRNA made than in the empty vector control. Because I am looking for two proteins (so two constructs) I could use one protein-construct as control for the other but we don't know enough about the proteins so I think this isn't a good idea (they could maybe work together somehow or s.th. else).

So, is there a better control than emtpy vector? Some protein-overexpression that has no effect (if something exists)? Or is my empty vector control just fine? Or is the other construct as control a good idea?

Maybe there is no perfect control...
But I just wanted to adress this question because of the highly importance of good controls.

Thanks in advance

[Lastwish]

empty vector is fine,i think
or,maybe you can set a known protein linked with GFP as control....

[Grenouille]

thanks for your reply.
a protein with no (overexpressing)effects (like what?), or just any protein?