i read that 500micro litre of bacterial culture with glycerol 500 micro litre (30%)
then in freezer this method is true??
i attach image that show the method >>> if this method is true how long culture is preserved ???
Glycerol is a cryoprotectant, which is why it is used. You typically want 15-20% glycerol with your bacterial culture to effectively protect from ice crystals. You can use other cryoprotectants like DMSO, but glycerol is cheap and usually used for bacteria. I typically use sterile filtered 30% glycerol, and mix 1:1 with my overnight bacterial cultures because it is easier to handle- the suggestion above will work as well but I get annoyed by the viscosity. ;)
Keep the glycerol stock at -80C for best stability (unless you really want to use liquid nitrogen, which wouldn't hurt but isn't necessary). When ready to use you can use an inoculation loop to streak some of the glycerol stock on to a plate- be sure to keep the stock itself frozen and it will last a long time!
i havenot freezer with -80 degree ..... so i want to preserve in normal freezer ??
yes. those will still work. store in deep freezers. check once every month to make sure stocks r fine. i guess it shud not b a problem for 3-4 months...let see.
dr Jennifer Gibbons >>> how long bacterial culture can be preserved with this method?
i isolate the bacterium in its isolation medium (Halophilic nutrient broth) that i take 500 microlitre from this culture to glycerol in normal freezer (-5 degree approximately) .... this is ok dr or not ??
dr Sansrity Sinha
i have 300 bacterium and it is difficult to subculture all bacteria every month
I agree with Jennifer Gibbons answer and suggstions. Else, it should be considered to for further use to streak some of the amount on solid agar media and in appropriate liquid media and verify purity and culturability.
Yes, that is ok.
Since 2 year, I have preserved bacterial cultures in 40% final conc. of glycerol in deep freezer about -20 Celsius. It still work ok.
I agree with Jennifer. In addition in our laboratory, we make a slight modification due to some problem with electricity (inconstant freezer's temperature), so we centrifuge it and discharged supernatant first before adding 1:1 sterile 30% glycerol. And in our experience it viability good enough.
Regards
dr Heru - Pramono
so the tube contain contain pellet of the microorganism and known amount of glycerol??
If we grow bacteria on BHI, or any other media other than LB, can we still use LB broth with glycerol to store the grown bacteria?
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