[Microbiology] Re: doubt in reviving bacterial culture from glycerol stock

John via microbio%40net.bio.net (by yjgent from verizon.net)
Thu Jun 30 21:44:37 EST 2011

On Jun 25, 9:33 pm, Kanchanadevi k <kde... from gmail.com> wrote:
> Hi all
> I want to revive bacterial culture from glycerol (70%) stock stored in -20
> for two years
> I tried by inoculating glycerol stock in nutrient agar plate and nutrient
> broth.
> But, No growth has come
> so I couldn't revive that culture
> can anyone tell me
> what is other possible way to revive the culture from glycerol stock?
> Thank you so much
> --

What is the organism? Does it have any special growth requirements or
nutrients? You need to use the proper media and atmosphere and temp
that would allow the best chance for the organism to shake off the
effects of freezing. Sometimes holding the broth longer might allow
for a few hardy bugs to revive.
You might need to consider thioglycolate broth or laked blood broth
for some fastidious organisms. A candle jar can increase CO2 levels,
but a strict anaerobic organism might need an anaerobic chamber or
Do you know what the generation time is for this organism? A slow
grower like Mycobacterium may need to be held for several weeks and a
Mycobacterium specific broth like 7H11 would be needed.

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