When I prepare NGM plate for the growth of C. elegans, the plates are clear. However, when I spread culture of E. coli OP50 on NGM plates and incubate at 37 C overnight, I observed crystal like structures. These structures were never found on un-inoculated NGM plate. What are the possible reasons and how these structures can be avoided?
Did you take the picture of those crystals? If yes, then maybe you can share with us how it looks like. I never experienced such situation before. 1 possibility was maybe the apparatus you used to prepare the E. coli was not clean, maybe it had some crystals substances.. Perhaps you can try to use new apparatus when you prepare the E. coli.
Some of the salts used in your medium. Probably you incubate E. coli thus reducing the concentration of water in the gel, making some of the salts precipitate making mineral crystals.
I have had a similar issue. But mine is with or without the culture. Sometimes I get those crystals, sometimes not. I am not sure what's the reason.
That could be the case of the salts precipitating out. Make sure your NGM is under 55˚C before adding the CaCl2, MgSO4 and KPO4 buffer. We've also had good results with just letting the seeded plates stay at RT for a couple of days; no need for 37˚C incubation.
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