plasmid transfection - (Jul/10/2008 )
We are going to transfect 2 plasmid into a mammalian cell.
Now we are wondering about since these 2 plasmids are from the same vector (pcDNA3.1).
Will there be any problem simliar as in bacteria which exists plasmid incompatibility issue?
any suggestion help
There is no problem with transfecting cells with two plasmids of the same vector. You just need to know that not every cell is going to pick up both plasmids and you are going to have some cells only expressing one of the plasmids while other cells are only expressing the other plasmid. However, the vast majority of your expressing cells should be expressing both as long as the purity of each DNA prep is nearly the same. If one of your plasmids is dirty (ie: high protein, residual ethanol) then this plasmid will not be transfected nearly as well as the other. What are you planning on doing with these cells? If lysates, it's not quite as much of a concern but if you are doing IF you are going to have to show that your cells are expressing both proteins.