I'm trying to grow some E. coli strains in minimal medium containing free fatty acids. I've used Triton x-100 (1.5%) to disperse the fatty acids but bacteria didn't grow. I also tried adding the fatty acid in a minimum amount of ethanol but I still see the fatty acid floating on the surface. what can I do to disperse them?
Noura, sometimes ultra-sonication helps disperse insoluble material.
Thank you Prabodh. I just tried sonication, it didn't work too. Do you think 1% is too high?
Try to use Tween 20 as a surfactant ad different concentions starting from 0.1 %
Hi
Try glycerol and heating while agitating the fatty acid molecules in the aqueous phase containing glycerol (pretty much like formation of liposomes without using solvents by Heating Method, see the attached paper).
Use Tween 40 or Tween 80 and then apply sonification for 90 minutes in ultrasonic bath..
Thank you very much everyone for your kind answers. I'm trying to measure the effect of exogenous fatty acids so I can't use Tween 40, 60 or 80 for that experiment. I already used them in the preliminary experiment as sources of soluble fatty acids. I will definitely try the heating method.
Let me know how it goes. Remember: there is always room for "modification" of techniques.
if you can get fatty acid-free serum albumin, you can try to make a mixture with your fatty acids and then aplly to the bacteria culture.
Another way is to dissolve the fatty acid in a solvent such as ethanol or isopropanol, put some drops of the mixture on the bottom of your petry dishes,, dry the solvent under sterile nitrogen flux and then add your bacteria
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