Except lyophilization what is the best why for bacterial culture storage?
Making glycerol stock (10-20% glycerol) is best method to preserve bacterial culture.
HELLO
I would add equal volume of 50% glycerol to the bacteria culture media and store them at -20 degrees.
thanx all
by using glycerol if storing at -20 degrees is necessary?
Glycerol stock is the best method . Of course ,you have to store at -20 degrees.
I agree with all the previous answer glycerol 10-20% added to the bacterial culture well preserve it for so long time.
but also soil preservation after sterilization of soil particles pure bacterial cultures added and then stored at -4 or 0 degrees for long time.
Grow the culture in their respective media to mid log phase, harvest by centrifugation and then mix this in 50% glycerol (media + glycerol). leave on bench for couple of hrs and then store at -80 degree Celsius.
I used to grow the bacteria in TSB (tryptic soy broth) plus 15% glycerol for 24 hrs. After that, freeze at -20 or better at -70 Celsius degrees
Of course, for fastidious bacteria you may change TSB by blood. Other may need special media
What we usually do, and it's been working well so far, it's to make a cell suspension into 1 mL of LB broth (from a fresh LB plate of the strain you wanted, streaked the day before), and then make 1 mL aliquots consisting in 500uL of the cell suspension + 500uL of 100% Glycerol (which will become a final concentration of 50% Glycerol). Then I would store it at -80C as our stocks at -20C have always had problems in some regard, many of them died, ending up loosing strains, thank God we had a -80C sample of the same strains.
Of course before storing the vials in the freezer, they should get mixed homogenously by vortexing and also put into dry ice, so the freeze gradually without getting damaged.
You may also use a massive 24 h growth in TSA, MHA o BHIA. Harvest and make a suspension in 2 ml TSB, MHB, BHI + glycerol 15%, Final volume 2 ml in Eppendorf tubes. Frezze a -20 o better a -70 Celsius degrees
We usually mix our bacterial culture with a final concentration of 30% glicerol and keep them at -70ºC for years.
In our lab, most of bacteria are keeping in TSB + glycerol 15-20% at -70ºC.
Good luck
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