What is the best way to preserve non-sporulating bacteria for further study?
Hi! It definitely depends on the bacteria. If you are working with the easy ones such as Enterobacteriaceae, glycerol with trypticase soy broth and preservation at -20ºC or -80ºC is useful. However, fastidious bacteria such as Pasteurella, Haemophilus, Campylobacter, etc. may not survive in those conditions and it is better to use skim milk medium or beads, at -80ºC.
Dear Charles,
we store most of our strains (sporulating and non-sporulating) at -80°C in cryovials containing a complex medium (e.g. BHI or LB) and 15-30% glycerol.
Best,
Michael
For plate cultures, resuspend the bacteria in 15% glycerol by vortexing then let sit at RT for 5 min then place in -80C. For liquid cultures, add DMSO to 5%, mix then place at -80C. Most bacterial species will remain viable for at least 20 years.
If it is just short term and you do not have a -80 freezer, you can store at -20 for a year or so in a storage media (no defrost cycle). Flash freezing with liquid nitrogen first always helps.
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