Have a PSL lab technician or a graduate student show you how to use the autoclave before operating it. Use caution when working with the heated autoclave. Always wear autoclave gloves when handling hot materials out of the autoclave. Also, use caution when opening the autoclave door after the process is completed, because the steam may cause burns. Never open the autoclave door if psi is 0.4 or above.
20 minutes for autoclave to get ready; around one hour for autoclaving (exact time depends on the materials being autoclaved)
| Step | Image |
|
The autoclave is located in 1122 PSL. |
 |
|
Check to be sure the deionized water is on by turning on a deionized water faucet in the autoclave room. The deionized water faucet is the far right knob at the sink.
|
|
|
Turn on the autoclave-steamer. The steamer is located in the Graduate Student Office (1115 PSL), inside the west closet. If the office door is locked, ask a graduate student for a key. It will take about 20 minutes for it to get ready for use. |
 |
| Step | Image |
|
Make sure lids are loose-fitting. If they are screwed on tight, there is a possibility that they might explode. Stick a piece of autoclave tape on the lids. Always be sure that the containers and lids are autoclave safe; if unsure, look up the product in a catalog or ask an experienced person. Aluminium foil may be used in place of lids for sterilizing beakers or measuring spoons. |
 |
|
Place containers on the racks in the autoclave. |
 |
|
The autoclave must reach 20 psi before starting. |
 |
|
Set your steam sterilize time to 45 minutes. In addition, if you are concerned about condensation buildup inside the containers, set steam dry time to 20 minutes. |
 |
|
Close the door and be sure it locks in place. Locking will occur as the wheel is spun. Make sure that there is a tight seal on the door. |
 |
|
Press the gravity button to start the machine. |
 |
|
If steam is escaping from the door, turn the machine off by hitting the white reset button. Untighten the door, push in on the door, and retighten it.
|
|
|
Fill out the sign-in sheet in the autoclave room.
|
|
|
The autoclave alarm will go off when the door can be opened (a slightly longer time than the run time you set). Be careful when opening the door, as steam may come out.
|
|
|
Dark stripes will appear on tape, confirming that materials have been sterilized. |
 |
| Step | Image |
|
As with a dry run, make sure lids are loose-fitting. This allows liquid to escape the bottle as it boils during the sterilizing process. Since liquids will boil in the autoclave, place bottles in a pyrex dish from the lab. Fill the dish with one inch of water. Stick autoclave tape on the lid. |
 |
|
Set your steam sterilize time according to the volume of liquid you are sterilizing. (Refer to the reference table below for some common values, or to pg. 155 of Phytotechnology Laboratory's catalog for other recommended autoclave times and volumes.) Do not set a steam dry time. Double-check that the time is set for zero. |
|
|
Close the door and be sure it locks in place. Locking will occur as the wheel is spun. Make sure that there is a tight seal on the door. |
|
|
Press the liquids button to start the machine.
|
 |
|
Once the autoclave has been started, follow the dry-run procedures for checking the door seal and removing materials.
|
|
|
Allow liquids to cool for 15 minutes before tightening lids.
|
|
Steam Sterilize Times | |
|
Volume of medium per |
Minimum autoclaving |
|
250 |
31 |
|
500 |
35 |
|
1000 |
40 |