Autoclave (OGL)

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The protocol for using the autoclave in OGL.

Overview

An autoclave chamber is used to sterilize laboratory equipment, including liquids, by subjecting them to steam created at a high temperature and pressure. Autoclaving should be performed on the following items:

  • Containers used while working with microorganisms

  • Glassware used to store food for experimental organisms

  • Biohazard waste containers

  • Pipette tip boxes

Tips before starting

  • Review or take the autoclave training on the Northeastern Environmental Health and Safety website.

  • It is mandatory to scheudle a 20 minute in-person training with Rosie Falco Poulin (r.falco@northeastern.edu) at the MSC to follow up on the online course.

  • Always unscrew jars or vials as the high pressure may cause them to explode if tightened.

Protocol

  1. Fill out autoclave sign up in binder

  2. Make sure all knobs are at zero

  3. Load samples onto metal sheet inside the autoclave

  4. Reset red arm to zero (top gauge). This arm keeps track of temperature maximum during the run

  5. Turn ON autoclave with green switch

  6. Turn bottom knob to fill water - wait until water reaches the front of the groove (might already have water in it)

  7. Switch knob to sterilize

  8. Seal the door by turning clockwise (not too tight)

  9. Look on the chart:

    • Determine what you are autoclaving (wrapped vs unwrapped)

    • Set temperature to the desired setting

    • Example: Unwrapped = 3850 model, cold start, 32 min, 134 degrees

  10. Set timer (NOTE: Setting timer starts the run)

  11. Come back 52 minutes later (32 in autoclave + 20 to depressurize) - If the red gauge reads lower than your desired temperature, it didn’t get hot enough (due to too much water or too much glassware). Run it again, if needed, for slightly longer (~40 mins) and make sure it reaches the appropriate temperature (indicated by red arm)

  12. Crack open door to boil off water

  13. OPTIONAL: Set bottom knob to extra dry (if there is still a lot of moisture

  14. Run for 20-30 min on dry

  15. Otherwise, set knobs back to zero and turn off autoclave

  16. Make sure the red arm is back to 0