Creating Stock Cultures of Bacteria

Creating Stock Cultures of Bacteria

1. Heat an appropriate media (usually complex like LB, 100 mL) up to 37°C in an incubator.
2. Inoculate the media with bacteria. This is usually done from a colony on a plate or from a stock culture.
3. Start shaking the media in the incubator after inoculation.
4. Allow the culture to grow to a visibly high density (4-8 hours).
5. Grab a bunch of 1-2mL microcentrifuge tubes and glycerol.
6. Pipet 0.5 mL of the dense culture into each microcentrifuge tube.
7. Add glycerol to each tube (30-50%) and vortex until thoroughly mixed.
8. Place all tubes into the -80°C freezer for long term storage.