In article <Pine.3.87.9410041720.A23832-0100000 at raven.csrv.uidaho.edu>,
jablonsk at RAVEN.CSRV.UIDAHO.EDU (Peter Jablonski) wrote:
> Does anyone have any information of the lyophilization of alkaliphilic
> halophiles or halophiles in general? We are trying to lyophilize
> halophiles for the purpose of storage, but haven't come up with a
> suitable medium to wash the cells after harvest.
The protocol below is used by the Australian Collection of Antarctic
Microorganisms (ACAM) for all their strains including extreme halophiles
(Halobacterium spp). This avoids the need to suspend cells in anything
other than their normal growth media.
Prepare a 10% (wt/vol) skim milk in distilled water solution. Dispense
0.2mL into prepared ampoules, seal with cotton plug and sterilise at 108C
for 30 min. When cool, freeze then freeze-dry. Aseptically add 1 drop only
of cells suspended in growth media directly onto freeze-dried milk pellet.
Partially constrict ampoule in flame. Place on manifold on freeze-drier.
Freeze-dry for 5 hrs or overnight. Seal ampoules with flame.
Hope this helps.
Kev
--
Kevin Sanderson
Department of Pathology email: Kevin.Sanderson at path.utas.edu.au
University of Tasmania phone: 61 002 35 4825
