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Adding glycerol to nutrient broth culture?


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#1 rocksan

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Posted 24 March 2009 - 12:03 AM

Hi all,

Im doing PCR, my TAQ polymerase just ran out, and my my newest order should arrive in some time. Im havin time constraints, and this is a last minute problem. X(

Is there a known method which i could use to preserve my bacteria in Ramsay broth already made just by adding some amount of glycerol into the broth?

Else i have to redo the bacterial broth to get a fresh viable one for DNA extraction..

BTW, my broth has 9970uL Ramsay broth, 20uL MgS04 and and 10uL Glucose. So 10uL.

Thabks in advance 4 anyone who can help me here..

Edited by rocksan, 24 March 2009 - 12:05 AM.


#2 molgen

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Posted 24 March 2009 - 12:36 AM

I use 15% glycerol (final concentration) to preserve the bugs.

But, if you want to do PCR on them, why donít you pellet them and then freeze them until you get the TAQ?

#3 pito

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Posted 24 March 2009 - 07:23 AM

I use 15% glycerol (final concentration) to preserve the bugs.

But, if you want to do PCR on them, why donít you pellet them and then freeze them until you get the TAQ?



@molgen:You simply add glycerol and store them on room temperature or ?




In general (so not speaking about using your broth) I know of glycerol being used as a protectant when freezedrying micro-organisms or when storing them in liquid nitrogen or sometimes simply in the freezer or fridge.

I did store a fungi using a simple water -glycerol (I used 10 up to 30% glycerol, but cant remember at the moment what gave the best results and I do not have my papers here) mixture and placed them in the fridge or freezer. Both didnt work really good, but I could preserve the fungi for 1-3 months.
Keep in mind that in the fridge it can happen that it continues to grow (slow but still) and that the micro-organism can stay long in a small tube, so dont keep it too long in there.


PS. its possible to preserve micro-organisms in simple water so I wonder how long you need to preserve them?
You can simply preserve the bacteria in water... for a few weeks, months and some even up to more then a year.

Edited by pito, 24 March 2009 - 07:40 AM.

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