Which cells? E.coli can stand -20 with 10% Glycerol but better at -80 for periods longer than 6 months. In general you have to pellet down the cells by centrifugation at low speed (5000g) then discard the super and resuspend the pellet with LB and Glycerol or PBS and glycerol.
Alessio
DIYbio Groningen
Sent from my iPad
Sent from my iPad
Hey--I wanted to try out freezing cells in a -20 freezer at 10% glycerol a few days ago. So I did it, and it looks like they are frozen solid. I don't know much about freezing cells, but I assume since they are solid they should stay good for longer. I forgot to check the ones with 40% glycerol, but perhaps they aren't frozenHas anyone tried this? Does anyone know if cells that are frozen solid will degrade over a long period of time? Of course I am trying it, I have cells in 10% 20% and 40% glycerol in both -20 and -80, but I'd like to know if anyone has experience with this. In a month or so I'll try to recover some plasmid from them. I'll just keep trying each couple of months and if it's working I'll start doing it ever 3 months. I am putting it directly into liquid culture, so all I am testing is if there is still plasmid, so I don't know how useful the results will beAnyone have any experience with this? (BTW NEB says that their comp cells become less competent in -20, but did they freeze them solid? Is there a difference? Sorry, I am no expert at this, so that's why I am trying it)-Koeng
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