Hi Sebastian,
I use one part bacterial culture and one part sterile glycerol. Mix thouroughly with a wide bore pipette (or chop the end off a 1000 ml blue tip.) They last at least a year at -20oC, maybe longer. (E coli-based clones).
Note: An agar stab in an eppy with a gas permeable top and stored in the fridge at 4oC lasts a few months. I use that method when I have a larger queue of clones than I can deal with right away but don't need long term storage. Hint: Gas permeable bandages with sticky on the back make a great topper for these. Just cut it up, stick on, and fold the edges over. If you have a lot of clones use a 96 well plate and a whole bandage.
Good luck!
Stacy
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