E. coli preservation

Michael Witty mw132 at mole.bio.cam.ac.uk
Mon Jun 25 13:54:20 EST 2001


Dear Jim,
        you can't beat glass and a really good screw cap, in my opinion.
Mike.

On Mon, 25 Jun 2001, Jim Kami wrote:

> Mike
>
>      I was planing on sealing the plates with either that fancy sealing tape
> from one of the biotech supply houses or 3" wide packing tape from OfficeMax or
> Staples. I would like to avoid oxygen as it is my understanding that the success
> of stab cultures is due in part to the anaerobic conditions the cells experience
> when inbeded in the agar. This is also why I'm interested in this MgSO4 method
> (unless someone was shining me on)
>
> Jim
>
> Michael Witty wrote:
>
> > Dear Jim,
> >          this sounds like a good experiment.  Are your 96 well plates
> > permeable to oxygen?  Why dont you try 10 or so glass containers too, to
> > that the effect of exposure to oxygen can be tested.  I have a
> > suspicion...
> >
> > Mike.
> >
> > On Sun, 24 Jun 2001, Jim Kami wrote:
> >
> > > Hello again,
> > >
> > >     Base on what I've been reading, it seems stab cultures are the most
> > > popular method for storing bugs so I'm going to try an experiment using 96
> > > well plates filled with LB + low melt agarose (so I can use a warmed
> > > autoplate filler to save time) and inoculate them with a 96 pin replicator
> > > tool and seal with tape. Let you know how it works in a year or two.
> > >     In the mean time, I recall being told many years ago, that storage
> > > under high molar MgSO4 is equally effective in preserving bugs
> > > "indefinitely". Has anyone heard of this method and know exactly what
> > > concentration of MgSO4 to use ??
> > >
> > > Jim Kami
> > > Rm 272 Hunt Hall
> > > Dept. of Agronomy
> > > University of California
> > > Davis, CA   95616
> > >
> > > Chris Fields wrote:
> > >
> > > > Tim Spahlinger wrote:
> > > > >
> > > > > Not to belabor the point, but I just happen to have found
> > > > > Sambrook's and Russell's procedures for Storage of Bacterial
> > > > > Cultures in "Molecular Cloning", Third Edition, page A8.5
> > > > > (appendix 8).  (I also needed to do some storage, so I
> > > > > looked them up!).  They don't give a relative time limit for
> > > > > the storage of stab cultures.
> > > > >
> > > > > By their methods, a final concentration (v/v) of 15%
> > > > > glycerol seems sufficient for long term storage of the bugs.
> > > >
> > > > I think our stabs were 1/2 conc. nutrient agar in plain storage vials,
> > > > stored in the dark.  The last bit, about storing in the dark, is a bit
> > > > baffling.  I was told that it is b/c of possible accumulations in
> > > > mutations under low-level UV (I think this is crap, but I'm also a
> > > > paranoid grad. student).
> > > >
> > > > --
> > > > C. J. Fields
> > > > Graduate Student, Dept. of Biological Sciences
> > > > The University of North Texas
> > > > Denton, TX
> > > >
> > > > email : cjfields at jove.acs.unt.edu
> > > > \\\\\\\\\\\\\\\\\\\\oooooooooooooooo////////////////////
> > > > "Giving money and power to government is like giving
> > > >  whiskey and car keys to teenage boys"
> > > >                                 -P. J. O'Rourke
> > > > "Join the military.  Travel to exotic places, meet
> > > > exciting people, then kill them"
> > > >                                 -Anonymous
> > > > ////////////////////oooooooooooooooo\\\\\\\\\\\\\\\\\\\\
> > >
> > >
>
>




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