Bacterial Glycerol Stock
- Agar plate with isolated colonies
- LB media
- Sterile flasks with ventilating caps or 50 ml tubes
- Screw-cap freezer vials
- Shaker incubator
- 5mL serological pipettes
- Micropipette and sterile tips
- Ethanol-dry ice bath
- -80oC freezer
- Fill 5mL media each into the flask/tube, aseptically.
- Add antibiotics if needed.
- Pick a single colony from agar plate and disperse into the media.
- Grow overnight culture of bacteria at 37oC and 300rpm.
- Add sterile glycerol (autoclaved) to 15% v/v final.
- Vortex vigoriously and dispense 1ml aloiquots in screw-cap freezer vials.
- Quick freeze on Ethanol-dry ice bath.
- Transfer to -80oC freezer for long term storage.