Another cryo problem

Rudolf Eichenlaub microbio at hrz.uni-bielefeld.de
Thu Sep 29 06:07:56 EST 1994


In article <1994Sep25.142848.8737 at mbcf> heath at mbcf.stjude.org (Richard) writes:
>In article <4ACCAD2D17 at botn.canterbury.ac.nz>, LYTTLE at BOTN.CANTERBURY.AC.NZ writes:
>>      Dear all,
>>      The comments over the last week or so about cryopreservation 
>> of microorganisms have prompted me to put my own problem before the 
>> net.
>>      I am working with four Comamonadaceae able to degrade 
>> in pure culture the nitrogen heterocyclic compound isoquinoline as 
>> sole 
>> carbon, nitrogen and energy source. Unfortunately they have a nasty 
>> tendency to lose the phenotype, especially when grown 
>> in a complex medium such as nutrient broth, LB medium and so forth. 
>> My problem is that to restore the cultures from stock at -80 C the 
>> organisms currently have to be grown in a complex medium or they won't 
>> provide a large enough inoculum for study.
>>       Does anyone know of a method of cryopreservation and 
>> restoration that may get round this? At the moment I am using the 
>> "Protect" bacterial preservers from Technical Service Consultants in 
>> the UK which use beads and a cryopreservative fluid. Any suggestions 
>> would be leapt upon with great glee and gusto!! Thanx in advance.
>> 
>> Trevor C Lyttle
>> Plant & Microbial Sciences Dept
>> University of Canterbury
>> Christchurch 
>> New Zealand
>
Have you tried to store your bugs as a water supension in the fridge?
Yust a thought. We always store our bugs (isolated from soil, probably
Commamonas) for up to one year now this way and they are much more viable
 compared to freezed cultures. If you make up your suspension from
a culture grown in minimal medium , you should be able to directly inoculate
cultures. This should reduce the probability that degradation genes
(perhaps due to an unstable plasmid) are lost. BTW, which medium do you
use? We can grow our bugs in liquid minimal medium, but not on MM-plates :-(

good luck, 
Anne


A. Schmitz                   email:aschmitz at biogene4.uni-bielefeld.de
University of Bieleled
Dpartment Gentechnology/Microbiology
Germany 



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