temperature for growing transformed bacteria - (Aug/19/2006 )

Hi,

I transformed chemically competent E. coli with my plasmid. After picking colonies, I cultured them overnight in LB medium. I was using a public incubator/shaker. I set the temperature at 37 degree. But apparently after I left, someone changed the temperature to 30 degree. This morning I checked the bacteria. The solution should be cloudy due to the growth of bacteria, but instead, it is still very clear. I don't know if it is due to the change of temperature. If yes, may I just change back the temperature and wait for another 24 hours?

Thanks!

hi,
did u add antibiotics?
at 30C also u shud see some growth ,
do u find any kind of thread like structures in ur tubes, if s then i can say may be ur cells got infected with phage.

with regards
payeli.

yes, I added ampicillin. I did not observe thread like structures.

Strange, overnight at 30°C is really fine (for thawn bacteria). I even found it better than 37°C because at 37°C overnight the cells tend to be at the end of the exponential phase, and there is a delay in the growth when I passage them. Personnallly I prefer 30°C overnight (for E. coli growing fast).
I don't think that 30°C is the reason of your problem.

But sometime you need a little more time for picked colony. let them in the incubator at 37°C, it will not hurt.

If you want to pick again some colony, I would first let the plate at 37°C for few hours (2-4 hours) before to pick colonies.

I don't think that 30°C is the reason of your problem.