In my experience that much glycerol at -20 will not freeze solid instead it is just a very viscus solution. I had a plasmid preserved in DH5's for 3 years that I had mistakenly placed in the -20 rather then -80 and they grew just fine when used to inoculate a culture. The only problem was that most of the cells had settled to the bottom of the tube.
It is my understanding that at -80 with a 40% solution storage is nearly indefinite with a cell line let DH5's
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On Fri, Jun 27, 2014 at 6:06 PM, Koeng <koeng101@gmail.com> wrote:
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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Richard Kramer
Cell and Molecular Biology
St. Cloud State University
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