Mike Barnkob:Protocols/Bacterial/Glycerol stock

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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


  1. After liquid culture has grown overnight, add 500 μL to 500μL of 50% glycerol in a 2 mL cryovial.
  2. Freeze the glycerol stock tube at -80°C. Should now be stable for many years.
  3. 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.