Autoclaving Laboratory Glassware and Media

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Autoclaving Laboratory Glassware (and Media)

Glassware that is to be used for cell growth must be sterilized beforehand. This is accomplished by first chemically disinfecting the glassware with 10% bleach or 70% ethanol (See wiki Chemical Disinfection of Glassware). Next, follow the procedure below for autoclaving.

  1. Collect all glassware that will be used in the upcoming experiment. Be sure to also collect any glassware that will be indirectly used: graduated cylinders, glass syringes, etc.
  2. If cell growth media is being autoclaved concurrently, make sure all glassware has a small amount of water (5-10% full) inside. If media is not being sterilized the water is not needed.
  3. Cover all openings of the glassware with aluminum foil. Use enough to securely hold the foil on the glass.
  4. Place into an autoclavable container. Polypropylene (PP) is a safe plastic that can withstand the autoclave. Be careful to not use polyethylene (PE), as it will melt in the autoclave! If autoclaving media, place a small amount of water (just enough to cover the bottom) in the plastic container.
  5. Use a cart to transport the container and glassware to the autoclave.
  6. Open the autoclave by holding the white button while gently pulling down on the door handle.
  7. Choose an appropriate autoclave program. 1 through 3 are LIQUIDS which are used to sterilize liquid media. Choose 4, which is VACUUM, if the glassware is empty.
  8. Sign the log book.
  9. Remove the container and glassware when the program is finished (~50 min). Be careful of the steam released from the autoclave when it is opened after being recently used. Also wear heat resistant gloves to pick up the container.
  10. Close the autoclave between uses. The autoclave needs to stay at about 115C to operate quickly.
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