AU Biomaterials Design Lab:Protocols/Autoclave

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An autoclave is a piece of equipment that is used for sterilization. In this lab, we primarily use the autoclave to prepare broth and flasks for bacterial protein expression. The autoclave that we use is located in Beeghly 314. There are directions posted on the wall in this room for proper use of the autoclave. We follow the directions for "Wet Samples".


  • Make certain that there is enough water in the reservoir before you begin.
  1. Turn the power on
  2. Turn the black knob to fill. This fills the chamber with water.
  3. Let water continue to fill the chamber until it reaches the line at the front of the chamber.
  4. Turn the black knob to STE (short for "sterilize").
  5. Load your media into the autoclave chamber. (Note: 2 2.8L Fernbach Flasks can be loaded into the chamber)
  6. Close the autoclave door and seal it shut.
  7. Set the temperature knob just to the right of 121F.
  8. Turn the timer knob to 50 minutes.
  9. After the 50 minutes have elapsed, wait 30 minutes until the pressure drops back to atmospheric pressure.
  10. Turn the black knob to Off.
  11. Use gloves to unload your sterilized material.