Adding glycerol to nutrient broth culture? - (Mar/24/2009 )

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..

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

But, if you want to do PCR on them, why don’t you pellet them and then freeze them until you get the TAQ?

molgen on Mar 24 2009, 09:36 AM said:

@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.