Any recommendations for very-low-fluorescence, rich growth media for E. coli? Teknova EZ Rich (or equivalent) seems to be the standard, but it's extremely expensive from our distributor and impractical to make from individual components at the small scale I need.
Neidhardt MOPS rich is the way to go. Life in research is always impractical... That said, you could try and cheat using a small amount of yeast extract and casamino acids to sub for all the other components in EZ rich. Casamino acids is poor in tryptophan, btw.
In our lab, we grow cultures for fluorescence microscopy in minimal medium supplemented with glucose and casamino acids, there is only very minimal background if at all. That said, if we needed a richer medium, we use regular media (like LB), after growth wash cells well (centrifuging at low speed) and resuspend in minimal medium before fluorescence experiment. Not sure about your exact application but hope this is helpful.
Thanks for your advice! I am doing long timelapse experiments, so washing isn't an option here. I usually use M9 minimal media with glucose and 1X MEM amino acids; but I'm looking for rich/defined/low-fluorescence media to get higher-growth-rate data. Supplementing with diluted yeast extract is a good idea for some preliminary experiments.
Follow-up/notes: I've been using EZ-RICH per the recommendations above. I see some precipitate in Teknova EZ-RICH if kept at 4 C or room temperature for more than a day or so after preparing it, so I keep 10 mL aliquots frozen and thaw as needed. Vortexing the media can break up aggregates and it doesn't seem to affect cell growth. I've also tried supplementing M9 media with EZ-RICH; a mixture of 99% M9 and 1% EZ-RICH results in growth that's nearly as fast as 100% EZ-RICH (probably not to as high density, but I haven't tested this).
You can try some of the defined media for protein expression from the Studier's paper: Article Studier, F.W. Protein production by auto-induction in high d...
.For example, MDG, LSG, P and their variations (Table 1).
"Fully defined media have been formulated with well metabolized amino acids at concentrations high enough to achieve saturation densities equal to or greater than those obtained in complex media. However, yeast extract appears to supply something that allows slightly more rapid initial growth than in those fully defined media."
I would expect them to give a decent E. coli growth and protein expression for most purposes.
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