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Making bacteria stock using glycerol


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#1 Nebraskagirl

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Posted 24 May 2015 - 02:25 PM

Okay I'm really new at this, I kind of inherited a lab with some issue.  My main problem right now is finding the freeze down protocol for bacteria.  From what I understand they are growing up the bacteria in about 8ml broth and adding 8 ml glycerol to this.  Mixing and putting into 1ml cryo tubes.  Somehow this doesn't seem right.  I used to freeze down virus in DMSO so maybe I'm wrong.  Could you some direction.

 

Thanks



#2 r.rosati

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Posted 24 May 2015 - 02:30 PM

You mean adding 8 ml of a 50% glycerol solution? Then it would be ok.

Would you like to check Addgene's protocol? It's really simple though. Bacteria are quite hardy (lyophilization is a valid preservation technique).



#3 Nebraskagirl

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Posted 26 May 2015 - 11:32 AM

I wish, no it seems they are taking it straight from the bottle and adding it to the bacterial broth.



#4 pito

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Posted 26 May 2015 - 11:44 PM

I wish, no it seems they are taking it straight from the bottle and adding it to the bacterial broth.

you mean they take the glycerol from a glass, commercial bottle?

 

Its not a good idea to just take stuff from a bottle (general solution), they should take aliquots or something.

In terms of concentration: if this is the right concentration.. then you can do it.


If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#5 Nebraskagirl

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Posted 27 May 2015 - 01:05 PM

Yes, no its not the right concentration.  I contacted the main branch of the school for their protocol and let's just say that I have my work cut out for me this summer.

 

Thank you guys






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