Revival of Freeze-dried Bacterial Cultures from DMSZ

The following protocol is used to reactivate and culture bacterial strains received from DMSZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH).

1. Remove the glass ampoule from plastic tube.
2. Check the indicator to ensure an intact seal (red or blue – good seal, pink or orange – humidity is present in the sample).
3. Heat the tip (pointy tip up) of glass ampoule over a Bunsen burner flame thoroughly.
4. Drop some water onto the tip to crack glass all around.
5. Carefully strike the glass tip with forceps (or a similar tool) to break off the tip.
6. Remove the insulation material and extract the smaller vial with forceps. If the bacteria in the vial is an anaerobic strain, ensure anaerobic conditions are used during the rest of the procedure.
7. Remove the cotton plug from vial and keep it sterile.
8. Add 0.5 mL recommended medium as indicated by the accompanying instructions to the dehydrated culture and insert cotton plug back into the vial. Allow the culture to rehydrate for about 30 min.
9. Remove the cotton plug again. Mix very gently with an inoculation loop and transfer to 5mL of a recommended liquid medium in a 15mL tube. Mix well.
10. Pipette 100uL of liquid culture onto plate as a spot and make a streak plate.

• Additional incubation directions are provided on the DSMZ website.