Normally, ecoli miniprep cultures are grown for around 12-18 hours at 37C.
However, due to my schedule, I won't be able to harvest the cultures for >24 hours. I think >24 hours is too long to grow ecoli cultures at 37C. I cannot freeze the cells and miniprep them later -- I will not be in lab at all for >24 hrs.
I want to grow them shaking at lower temperatures to slow their growth so I can harvest them later. However, I do not know what growth duration is appropriate at these temperatures.
How long should ecoli be grown at 30C and 25C to get something similar to 12-18 hrs at 37C?
It is totally safe to grow E. coli @ 30°C and 25°C for plasmids preparation. The time to reach culture saturation (stationary phase) will mainly depend on the initial load in your samples and its volume. In rich media supplemented with glucose, you should be able to get enough bacteria in a 12 hrs ON culture, in regular E. coli lab strains.
Hello Gen, how are you doing today?
Well, depends of E. coli competent cells. Some people reports that grow well at 30°C, but take more than 24 hours to achieve enough amount for harvest, being 37°C the optimal temperature for sure. I always used 37°C for bacteria growth. At 25°C may be your bacteria will not grow at all, is too cold for E. coli. Perform some trials at 30° and 32°C during 24 or 36 hours before stablish some protocol for you schedule. Make sure to use 37°C for SOC medium incubation to increase the efficiency of your transformed bacteria. Hoped helped and good luck in your research.
Hello Gen Gi ,
I guess it wont be a problematic. ideal temperature would be 37oC for E.coli. growth. If you are unable to harvest at 24 hrs. time it wont be more problematic. your yield will be increased.
You're playing with fire. I doubt anybody has conducted enough systematic study of this issue to know for sure what's safe and what isn't.
Growing them under substandard conditions can damage to your plasmid in ways that you don't necessarily see immediately. You'll end up investing time and work into a molecule that might turn out damaged in the end.
It's not worth the risk. Fix your schedule and use standard protocol.
It is totally safe to grow E. coli @ 30°C and 25°C for plasmids preparation. The time to reach culture saturation (stationary phase) will mainly depend on the initial load in your samples and its volume. In rich media supplemented with glucose, you should be able to get enough bacteria in a 12 hrs ON culture, in regular E. coli lab strains.
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