Background Info
https://ehs.princeton.edu/book/export/html/380
https://www.ehs.washington.edu/system/files/resources/autoclave-safety.pdf
The autoclave is used to sterilize glassware, metal tools, and liquids (growth media, water, solutions). It kills bacteria and spores through heat and pressure. Most items will be sterilized after 30 minutes at 121°C at 20 psi.
POTENTIAL HAZARDS
-heat and steam burns
-hot fluid scalds
-injury in case of explosion
-exposure to pathogens
PROTECTIVE WEAR
-goggles
-long sleeve pants and shirt
-closed toe shoes
PROTOCOL
Liquid Sample Prep
1. Fill container with liquid to be sterilized
2. Unscrew cap until loosely attached to bottle
3. Put a piece of autoclave tape on the cap
4. Use lab tape with end folded over to label bottle with name and date and contents
NOTES
-do not fill bottle more than half way because the liquid bubbles and will spill over
-do not autoclave an item with the lid closed, there is a risk of explosion
-never put volatile liquids or solvents or corrosive chemicals in the autoclave
Solid Sample Prep
1. Wrap item in tinfoil or cover openings with tinfoil
2. Place autoclave tape on foil
Prepare Autoclave
1. Close water outlet nozzle
2. Fill basin with 600ml distilled water
3. Add item to be sterilized
4. Screw lid on autoclave
5. Set timer to desired time (20-30m)
6. Set exhaust valve in up position to allow air to escape
7. Set temperature to correct position
NOTES
-do not touch side knob, just main knob
Run Autoclave
1. Turn on machine
2. Once steam starts spitting out of exhaust valve (after several minutes), close valves using gloves
3. Check that temperature gets up to 121C
4. Sterilizing light should turn on and timer should start going down
5. Once time is at zero the machine should beep
Stopping Autoclave
1. Turn off the power
2. Use a glove to open the steam
3. Once the pressure is at zero
4. Open the water outlet
5. Open the autoclave
6. Use gloves to remove item and close lid
7. Close lid on any liquid bottles and check that autoclave tape turned black
NOTES
-items will be hot, do not leave where they could burn someone
-let items cool to room temperature before moving to shelf
-do not autoclave plastics unless they explicitly say they can be autoclaved