We have at least 1 construct that a) works in mammalian cells and b) seems to be cloned properly by PCR digest, but that is growing very poorly in Stbl3 cells (10x less plasmid and lower OD than bacterial with the parent px330 plasmid). Has anyone experienced that before, or have a solution?
Hi Dudley,
I hope you are doing well, and everything goes well, too. I am using PX330 plasmid construct to generate a mouse model. When I discussed the optimized protocol with one of my colleagues at the Rockefeller, she definitely suggested me to use Top10 as the competent E.coli strain because they had bacterial growth issues in the past. Personally, I have never tried Stbl3 cells but Top10 comp cells are working well on my hands.
Best,
Yetis
Is there a particular reason that you need to use Stbl3 cells? They're helpful for preventing plasmid instability but they tend to grow slowly.
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