E. coli preservation

Chris Fields cjfields at jove.acs.unt.edu
Sun Jun 17 19:03:37 EST 2001

My professor has several strains of E. coli and Salmonella in stab
cultures that are still viable after 20 years!  I think that you have to
be VERY careful with the phenotype.  A few of his were DOA due to
thymineless death (had a thyA mutation).  I guess his grad student at
the time had left out the thymidine; my prof was a bit pissed.

Several of my E. coli and Pseudomonas strains in stabs have been fine
for the last three years with no detectable loss of viability.  I don't
store strains with plasmids this way (I freshly transform them) and I
always make sure to store additional stocks at -80deg and lyophilized. 
This way, if the stab dies, I have a failsafe.

I think Molecular Cloning by Sambrook has all of the procedures.

C. J. Fields

Nick Theodorakis wrote:
> In <001701c0f4b3$34b0cfe0$8c01a8c0 at tn.nic.in>, R. Jayakumar wrote:
> >
> >----- Original Message -----
> >From: "Nick Theodorakis" <nicholas_theodorakis at urmc.rochester.edu>
> >To: <methods at hgmp.mrc.ac.uk>
> >Sent: Wednesday, June 13, 2001 11:17 PM
> >Subject: Re: E. coli preservation
> >
> >
> >> In <00c401c0f42a$7238c220$8c01a8c0 at tn.nic.in>, R. Jayakumar
> wrote:
> >> >
> >> >Stab culture -- Mostly everybody would have thought of it,, but the
> >> >gentleman wanted to know something that would preserve his cells
> for
> >> years
> >> >and not a few weeks which stab cultures do.
> >>
> >> I used to routinely gets live bugs form 2-3  year old stab cultures.
> >>
> >> When I was a grad student, that was our labs preferred method of bug
> >> storage.
> >>
> >
> >
> >the more important question here is did you store them in room
> temperature.
> >If so, then your experience will be useful to me too..
> >  thanks in advance
> Yes. Room temperature, in the dark (a drawer). With a glass vial that you
> can close very tightly to keep the air out.
> Nick

C. J. Fields
Graduate Student, Dept. of Biological Sciences
The University of North Texas
Denton, TX 

email : cjfields at jove.acs.unt.edu
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