For reference: I am transforming a newly assembled plasmid
what happened: I accidentally incubated my electroporated e.coli cells on the shaker at 30C for 1 hr rather than 37C and did not realize until I plated my culture.
Should I still expect colonies? Or did I make a fatal mistake?
I am incubating over night at 37C and I have the rest of my culture that I did not plate at 4C so I may be able to use that?
Hi Dina, I usually transform recombinant plasmid to E. coli by heat shock. It's simple and convenient. For electroporation, My suggestion is continue to observe the growing colonies, 30C and 37C are different but you can still expect the colonies. If not, re-do the experiments. Personally, after transforming the cells should be plated right away on the plate, and no storing. You can try to place 4C-stored culture but I don't think it's a good idea.
Hi Dina, I think you should still have some colonies. You can try to keep the plate for longer than overnight if there are none.
E. coli survives & grows just fine at 30*C, it just grew a bit slower than it would at 37*C.
I'd say you can proceed with the plated cultures and you should get colonies like normal.
Just make a note of it in your lab notebook.
Dina good day. Surely you will get colonies, but a little later.
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