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E. coli (ATCC 25922) growth in Minimal Medium


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#1 Fiaq Khan

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Posted 26 June 2013 - 05:49 AM

Hi everyone,

I am trying to grow E.coli (ATCC 25922) in minimal medium of following composition (per liter): Na2HPO4 (68.16), K2HPO4 (3.83), NH4Cl (1.07), NaCl (0.50 g), CaCl2.2H2O (0.0147 g), MgSO4 (0.1 g) and glucose (4 g).

I tried to grow overnight culture in above minimal medium but bacteria showing no growth (OD600 does not show any growth).

Can somebody help me with this that why I am not getting any growth. Thanks

#2 phage434

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Posted 26 June 2013 - 06:34 AM

Check the pH of the medium. It should be around 7-7.5.
You could also try adding some of the common supplements, such as thiamine.
This is a wild type strain, and should grow in that medium.
Are you sure the cells are viable?

#3 Fiaq Khan

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Posted 27 June 2013 - 01:31 AM

Thanks for your reply.

The pH of the medium is around 7.4. I have not tried thiamine. Because I thought that it would grow without it.

How much thiamine should be used for optimum growth of bacteria and what is it function?

#4 phage434

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Posted 27 June 2013 - 05:40 AM

Where do your cells come from? How do you know they are viable?

#5 Fiaq Khan

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Posted 27 June 2013 - 09:40 PM

Where do your cells come from? How do you know they are viable?


I have taken them from Life and social science department. I have grown the cells in Nutrient Broth (NB) and they showed optimal growth. Overnight culture showing OD600 ~ 1.2. I also did viable counts and measure the CFU in NB showing approx. 2.8 x 108 cells/ml.

Edited by Fiaq Khan, 27 June 2013 - 09:41 PM.


#6 Phil Geis

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Posted 29 June 2013 - 05:48 AM

i assume you're running a parallel control in something like TSB.

I don't understand why folks are so reticent to look at the literature. The following reports a minimal medium supporting growth of the isolate you mentioned.
Antimicrob Chemother. 1992 Feb;29(2):121-7.

Maintenance requirements of Escherichia coli ATCC 25922 in the presence of sub-inhibitory concentrations of various antibiotics.




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