u/EquivalentEqual7315

Hi everyone!

I am using double nickase plasmid from santa cruz to knock out a gene. After transfection with the plasmids, I am selecting with puro for a day and sorting gfp positive cells into 96 well plate to grow single colonies.

Then I want to introduce another gene but available plasmids have only puro selection or gfp.

I was wondering if my KO will be puro resistant or gfp-positive. And if it is absolutely bad idea to try to introduce another gene.

I feel like double nickase plasmids are more transient and won't be stably integrated?