Will my strain in glycerol last? - 30% v/v glycerol doesn't hurt right? (May/27/2008 )

The amount of glycerol is beside the point. I'm maintained my strain as glycerol stocks at -20'C but when I use one tube as the working stock, I realised that I have sediments on the bottom of the tube. Looks like bacterial mass to me. Will my other stocks last? It'll be a nightmare if I were to lose this strain as I'll begin working on it in a few month's time.

Thanks all.

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You should be fine. I generally use 25% Glycerol (final conc) and store them at -80'c, but anywhere between 10 and 40% final glycerol is used, and stocks are kept anywhere between -20 and -80'C. Bacteria are hardier than mammalian cells. Just thaw them on ice.

Ideally, you should resuspend bacteria in Glycerol before freezing them down, so that Glycerol is equally distributed and the sedimentation would not occur, but if it did, no worries.

See some protocols here: http://search.vadlo.com/b/q?sn=158621799&a...stock&rel=0

If you are still worried, take out some representative vials, streak them on appropriate agar plates, don't loose your sleep over it.

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Alrite, could sleep better for the coming weeks. Thanks lots.

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By the way, I misspoke, like Obama or HRC, depending upon whether you are American and who you root for.

You don't really have to thaw them on ice, just stick or touch a probe in the frozen slab, and streak it on the plate.

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