Mike Barnkob:Protocols/Bacterial/Glycerol stock
How to create a glycerol stock.
Prepare 50% glycerol stock:
- Dilute 100% glycerol to 50% with dH20
- Add 500 μL of 50% glycerol stock to appropriate number of 2ml CryoVials
- After liquid culture has grown overnight, add 500 μL to 500μL of 50% glycerol in a 2 mL cryovial.
- Freeze the glycerol stock tube at -80°C. Should now be stable for many years.
- To recover: open the tube and use a sterile loop or pipette tip to scrape some of the frozen bacteria off of the top. Do not let the glycerol stock unthaw! Streak the bacteria onto a LB plate containing the appropriate antibiotic.
Note: Keep cryovial on dry ice when retrieving bacteria.