Koch Lab:Protocols/How to Autoclave

How to use the Autoclave

Before using the autoclave, make sure that there is enough water in the reservoir. To do this, pull out the dipstick on the right side of the autoclave and look on the inside half of the strip. The indicator will be wet, and there should be enough water to wet the stick somewhere between the "Min." and "Max." positions. If it is low, fill it up with some DI water, but be careful to not exceed the maximum volume.

Sterilizing the Samples

1. Mark everything that you are putting into the autoclave with the heat sensitive tape. The tape should be in the drawer under the autoclave.
2. Place everything on the tray with some room between each item. If you are sterilizing liquids, use the non-perforated pan.
3. Flip the green switch, upper right corner, to the "START" position.
4. Turn the bottom knob, clockwise, to the "FILL WATER" position.
5. Watch the water fill in on the inside of the cavity, when it reaches the front of the cavity, the small notch, turn the bottom knob to the "STE" position.
6. Close the door and tighten it.
7. Set the temperature knob, second from top, to your desired temperature.
   • 273°F for plastics and other solids.
   • 212°F for the liquids.
8. Set the time, top knob, to the desired time.
   • For plastics and solids 45 mins.
   • For liquids 60 mins.
   • NOTE: If the autoclave is already warmed up, subtract 10 mins from these times.
9. The orange heat light, the lowest light, will turn on and the autoclave will run on its own.
10. After the autoclave has ran through its cycle it will ding to inform you it is done. Depending on what you want to do follow one of the steps below.

Drying the Samples

1. Wait for the pressure sensor to register 0 psi pressure difference between then inside of the autoclave and the lab. This is indicated by the white pointer on the upper dial pointing to 0.
2. Slowly loosen the door about an inch to release the steam. Leave the door open to allow the steam to be released throughout the drying cycle.
3. Turn the bottom knob to "EXH + DRY"
4. Set the timer to ~20-30 mins.
5. To indicate that the autoclave is working, the orange dry light, the middle light, will come on.
6. After the cycle finishes, turn the bottom knob to the "0" position, open the door and remove samples.
7. Flip the green switch to stop when you are done.

Not Drying the Samples

1. Immediately turn the bottom knob to the "EXH + DRY" position.
2. Wait until the pressure drops to 0 psi.
3. Turn the bottom knob to the "0" position.
4. Open the door, and remove the samples.
5. Flip the green switch to stop when you are done.

Liquids

1. Leave the bottom knob on the "STE" position, and wait for the pressure to drop to down to 0 psi.
2. Turn the bottom knob, clockwise, to the "0" position.
3. Open the door and remove the samples from the autoclave.
4. Flip the green switch to stop when you are done.

REMEMBER: The autoclave is hot, very hot, so anything you put into it is also going to be hot when you are done with it. That being said, always use caution taking things out of the autoclave, you don't want to get burned, drop something or spill any liquids on you lap.

Cleaning the Autoclave

If you ever need to clean the autoclave, refer to this journal entry on the procedure on how to clean it. It is recommended by the manufacturer to clean the autoclave once a week, or every twenty uses, which ever one comes first.