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autoclave.md

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Autoclave use protocol

  1. Before autoclaving, make sure:
    • Everything being autoclaved has autoclave tape on it (the tape indicates if items have been properly sterilized) except for biohazardous waste bags
    • Any screw-on lids are loosely screwed onto any bottles going into the autoclave (if screwed down all the way, bottles could explode during the autoclaving process)
      • Exception: Glycerol tubes. Twist caps down most of the way, but not tightly. This is because a lot of liquid is lost if caps are loosely screwed down.
  2. Load items to autoclave into autoclave bin
    • Liquid and dry items are autoclaved seperately
    • 3 L of liquids max
    • Don't stack liquids, but you can stack dry items
  3. Place the autoclave bin into the autoclave
    • Biohazard bags shouldn't be touching the sides of the autoclave
  4. Shut autoclave door all the way
  5. Select the proper protocol from the menu
    • Press "Select cycle", scroll to correct protocol, press "enter" to view protocol
    • Note: Anything with agar in it needs to be autoclaved on a liquid cycle! This includes biohazardous waste with agar plates in it. If not autoclaved on liquid cycle, liquid will overflow into autoclave and cause expensive issues.
    • Liquids:
      • 500 mL: P13 liq LIQUIDS1 (15 min exposure time)
      • 1-1.5 L: P15 liq LIQUIDS1 (20 min exposure time)
      • 2-3 L: P14 liq LIQUIDS1 (30 min exposure time)
    • Dry: P8 grav GRAVITY1 (30 min exposure time)
    • Orange biohazardous waste: P14 Liq LIQUIDS1 (30 min exposure time)
    • White biohazardous waste: P8 grav GRAVITY1 (30 min exposure time)
  6. Press "Start" to start cycle
  7. When cycle is finished, autoclave will ring
  8. If autoclaving liquids, take items out of autoclave
    • Wear protective gloves and be careful—items will be very hot
    • If autoclaving media with agar for pouring plates, immediately put media in 55°C water bath
    • For other liquids, leave out at room temperature
  9. If autoclaving dry items, crack open the autoclave door about a couple inches and let items dry for a few hours or overnight, then take out of autoclave
    • Items will still be hot after drying, so wear protective gloves when handling the items
  10. If autoclaving liquids, immediately rinse out the autoclave bin with hot water to clean out any boil-over
  11. If autoclaving biohazardous waste:
  • If there is a lot of boil over, place autoclaved bag into another biohazard bag, dump the boiled over liquid into the bag as well, tie the bag closed for disposal
  • To dispose of autoclaved bags, put a green "OK to trash" sticker labeled with our lab name and room number on the bag, then place bag in big black trashcan in autoclave room
  • Make sure the big black trashcan isn't too full; if it's full, take the bag out, tie it closed, label with an "OK to trash" sticker, and set the bag to the side of the trashcan