Making bacteria stock using glycerol - (May/24/2015 )

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

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).

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

Nebraskagirl on Tue May 26 19:32:09 2015 said:

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.

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