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glycerol stock


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#1 chicken

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Posted 14 July 2009 - 08:36 PM

i have some lb broth for about 4 to 5 days old in fridge 4 c.

can i make there glycerol stock and also can i use them for growing new cultures.

confusion. how long a cultre can be placed in fridge 4c without glycerol.

#2 tsng

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Posted 14 July 2009 - 09:06 PM

I've kept cells for a month without glycerol, but obviously not advisable as most cells die once it runs out of nutrients.

You can use your glycerol stock to make a new stock. All you really need is to scrape some off your glycerol cell stock with a heated loop or pipette tip and put into LB media.

#3 leelee

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Posted 14 July 2009 - 09:54 PM

Take some of your refrigerated culture and culture for several hours, then make your glycerol stock from that.
It was my understanding that glycerol is added to a culture prior to freezing to help prevent ice crystal formation, therefore there is no need to have glycerol added to refrigerated stocks, however I do agree with tsng that long term storage of bacteria at 4C is unadvisable.

#4 HomeBrew

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Posted 15 July 2009 - 05:04 AM

i have some lb broth for about 4 to 5 days old in fridge 4 c.

can i make there glycerol stock and also can i use them for growing new cultures.

confusion. how long a cultre can be placed in fridge 4c without glycerol.


I agree with the others. There is no need for glycerol when storing at 4C -- glycerol is a cryoprotectant, and therefore is useful when freezing cultures, not when refrigerating them.

A good number of your cells at 4C will likely still be alive. Just swap the order of your intentions -- use the 4C cultures to start new ones, and make glycerol stocks from these new cultures.

As for how long a culture can retain viable cells at 4C, it depends greatly on the strain of bacteria, the media they're in, the density of the culture when stored, etc. But such a culture should retain some viability for a few days at least, several weeks at best.




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