co-electroporation - (Oct/27/2005 )
What is co-electroporation and when is it used? Are there any advantages over normal electroporation?
I read an article that pretended to co-electroporate a plasmid with their gen of interest and another plasmid with their resistance-gene. Why don't they just put them together in one single plasmid? They have no way to determine if the 2 plasmids were taken up.
Hi there,
each plasmid must have a different reporter gene for that to work (like AmpR and ChlR). Another possibility (was that an yeast article?) is that they are not co-transforming two plasmids but a linearized plasmid (preferentially with a gap) plus a DNA fragment (gene of interest) containing some homology with the linearized plasmid. The yeast cells will use homologous recombination and "clone" the gene for you.
Clarice
I read an article that pretended to co-electroporate a plasmid with their gen of interest and another plasmid with their resistance-gene. Why don't they just put them together in one single plasmid? They have no way to determine if the 2 plasmids were taken up.
I read an article that pretended to co-electroporate a plasmid with their gen of interest and another plasmid with their resistance-gene. Why don't they just put them together in one single plasmid? They have no way to determine if the 2 plasmids were taken up.
Co-transfection (or co-electroporation) is used when your plasmid doesn't contain a (usefull) resistance gene. If you co-transfect the plasmid of interest with 10 times more molecules then the plasmid with the resistance gene, you can calculate the probability that a cell that becomes resistant, did not take up your plasmid of interest. It is very low! You should also take into account that a cell that takes up plasmid DNA, takes a lot of plasmid molecules.