[Microbiology] Re: Microbio Digest, Vol 49, Issue 5

Rachna Singh via microbio%40net.bio.net (by rachnasingh.25 from gmail.com)
Sat Jun 27 21:28:57 EST 2009


 Re: glycerol stock for bacteria and fungi,
In our lab, we generally prepare glycerol stocks (bacteria) in brain-heart
infusion (BHI) broth with 15 % glycerol and have stored the stocks at -20 as
well as -80. Even we don't have access to -80 much. So, I generally use -20
stocks only. My -20 stocks, stored 3 years earlier, are working well - the
bacteria are viable. Though the recommended temperature is -70 to -80, you
can store at -20.

On Sat, Jun 27, 2009 at 10:35 PM, <microbio-request from oat.bio.indiana.edu>wrote:

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>   1. Re: glycerol stock for bacteria and fungi, (Bob)
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> ----------------------------------------------------------------------
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> Message: 1
> Date: Fri, 26 Jun 2009 16:03:06 -0700
> From: Bob <bbx107.XYZ from excite.XYZ.com>
> Subject: [Microbiology] Re: glycerol stock for bacteria and fungi,
> To: microbio from net.bio.net
> Message-ID: <jska459cou2kriq2k4o3gapsquog96l4bf from 4ax.com>
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>
> On Mon, 22 Jun 2009 04:34:33 -0700 (PDT), mohamed eda
> <medanrc from yahoo.com> wrote:
>
> >Dear all hope you all fine and doing well
> >I want to prepare a glycerol stock for bacteria and fungi, and I want to
> know if there are different techniques  for each,
>
>
> It is mainly empirical. If you have a procedure someone says works,
> try it. Otherwise, you might try using 20% glycerol, with good healthy
> cells.
>
> Run some tests to see if it works. Freeze, and thaw samples at
> intervals; check viability. If it is pretty stable over a week, you're
> probably ok.
>
>
> >the second point is I did not have a -80 degrees freezer only -30 which I
> have, so it is possible to use it for the storage of the glycerol stock of
> the isolates and in this case is there is any special precaution in case of
> using -30 degree freezer.
>
>
> There are two concerns.
>
> One is simply that lower T is better -- things are slower. Probably
> not a big deal.
>
> The other concern is whether the freezer holds T well. If samples are
> stored near  -20, T fluctuations can cause changes in ice structure;
> not so good.
>
> If -30 is what you have, then try it. But you might also look into
> maybe saving some samples elsewhere if  -80 or liquid nitrogen are
> available. Hey, storing some aliquots in at least a 2nd freezer is not
> bad anyway -- just in case something goes wrong.
>
>
> You might check with the ATCC. This is their business, and I suspect
> they would share some info with you.
>
> bob
>
>
>
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