All-
I am running an experiment for protein expression with e. coli and my LB plate behave strangely these days. As you see in the attached picture, even in several days after transformation, colonies start to behave like a 'old' one. (I have seen those behavior of colony which transformed more than a month.) Whenever I see that elongation behavior of colony to prepare my starting culture, I usually failed to protein expression in bacterial liquid media.
Does anyone know why those e. coli colony behavior happen?
Is that behavior related with dryness of e. coli plate?
Or does that usually happen in overgrown plate? (Or too much stayed in room temperature?)
Thanks.
Huihun: Did you include an antibiotics against which your vector carries the gene to it in your LB plate? You should get good colonies overnight and, perform an analysis, and then seal and move it to 4C for storage. Good luck.
Hi, Ke-Wei,
Sure. I am using LB/Kan50 plates. - for pET28b vector backbone with NEB T7 Express Competent cells. I transformed that plate on Oct. 28th but it looks it is old plate.
I do not know the reason, actually.
In my previous lab we had the same problem, and we found out that our LB plates were not good, so we needed new ones. If you had changes in temperature it may affect you LB plates efficiency.
Hi, Erika,
Thanks for the reply.
Actually, I am also suspicious on my LB/Kan50 plate. So I prepared a new LB/Kan plate and do a transformation again on a new plate to fix the problem.
Again, appreciated for sharing your experiences!
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