Common Autoclave Rules
1. Every lab was given trays – use them. All items to be sterilized, whether wet or dry, must be placed in a stainless steel tray. NO lab is exempt. Never place anything directly on the racks and never in plastic bins.
2. Don’t overfill biohazard bags. You should be able to close the bag. Use a rubber band or loosely tape the bag. Do not twist the bag, it won’t sterilize properly if the bag is sealed. Place the bag into the appropriate trash can immediately after autoclaving.
3. Do not place anything on the floor.
4. Pick your items up in a timely manner.
5. If the autoclaves are in use, take your stuff back to your lab.
6. Be sure that the autoclave area is left in an orderly condition. All debris must be properly disposed of and all glassware etc. removed in a timely manner.
7. Be patient. If the outside pressure is not at 15 lb. (see below), the cycle won’t start and you will get an error message.
Steris large and small autoclaves (NSC 528)
1. If you are the first user of the day, the large autoclave needs to have the outside chamber pressure up to a minimum of 15 lb and up to 21lb. Press RESET and come back in ~30 minutes.
2. Place items to be sterilized into a stainless steel tray.
3. Place trays into autoclave.
4. Close door and turn wheel until hard to turn. The display will indicate that the door is open, but even when the display goes away, continue tightening.
5. Select 1 -> 4. Select a # one time and you can see what the temp. and times are.
6. Once you know which #, press the # again. (Select 2 times).
7. Once inside chamber pressure is at 0, the display will tell you to “Warning, Hot liquids. Open door 1”; Wait 10 min.”
8. You can’t run the autoclave again until this 10 min. is up.
Amsco 3041-S (large autoclave, NSC 484)
1. Place stainless steel trays onto large rack.
2. Release rack by pulling small black lever to the left of the cart.
3. Release cart by pulling large black knob. Move cart away from the door.
4. Close door and tighten wheel.
5. Follow directions for Steris large and small autoclaves above.
Amsco SG-120 (small autoclave, NSC 484)
1. Place stainless steel trays onto rack.
2. Push rack into autoclave and move cart away from autoclave door.
3. Close door by using pedal near floor on the right.
4. Select cycle.
5. Select one of the cycles and the display will show the sterilization time and vent time for the run. Go to Previous if you want to look at another choice.
6. Start cycle.
7. Timer will show indicating how long the run will be. It’s pretty accurate.
8. Once your cycle is done, use the foot pedal to open the door.
Electric autoclaves
Sanyo Labo Autoclave (NSC 528)/New Brunswick AC-48 (NSC 484)
1. Warning: this autoclave takes a really long time to come up to temp., as long as 1 1/2 hrs. It is fairly common for someone to come in and not realize that it is in use and stop the sterilization and take your stuff out. Recommend putting a note on the sterilizer “IN USE”. If the “oC” is flashing, the sterilization is done and the pressure and temp. are coming down.
2. Use distilled H2O in autoclave.
3. Check to see if there is any H2O in the main chamber. To do this, wiggle the whole unit and look down in the chamber – if you see H2O through the round hole, do not add anymore H2O. There should not be standing H2O in the chamber
4. Remove the exhaust tank and fill with H2O to the High level mark.
5. Open the chamber door and pour H2O inside (~2 L) until you can physically see H2O in the round hole in the bottom of the chamber. This will take the H2O level in the exhaust tank to usually between the HIGH and LOW mark.
6. Fill the exhaust tank with H2O to the LOW mark and not over the HIGH mark. Connect the exhaust hose and place the tank inside the unit.
7. Turn the power switch on (MAIN).
8. The sterilization temperature (usually 121 oC) should not be changed. Set time, hold timer button down and use arrows to go up and down (there is a lag time for the sterilizer to reach the set temp. and this depends on the volume you are autoclaving; Example, for a 500 ml flask, there is a 15 min. lag, so for a 15 min. sterilization ,set the time for 30 min.).
9. Place the items to be sterilized into the chamber. There are 2 large round racks that fit into the chamber. Use the solid one for liquids. You can also stack the 2 racks if you have a lot to autoclave. Turn the exhaust knob (control panel right) fully clockwise until it becomes tight. (do not overtighten)
10. Close the chamber door. Although the door closure light indicator lamp lights up, continue to turn the handle until it becomes difficult to turn. Attach the magnetic safety connector to the back panel. The safety connector is usually wrapped around the lid; unwind and it clicks into the round connector on the left side panel below the main control panel. If for any reason this comes off, the autoclave will shut down.
11. Press the start button to begin automatic sterilization process.
12. Come back in ~20 min. to make sure that everything is going as you expect – correct time and temp.
13. After the sterilization process is completed and the chamber pressure returns to normal atmospheric pressure, the temperature inside the chamber will fall below 80oC. At this point the END buzzer will sound 5 X’s. Be sure that the gauge pressure reading falls below 0 before opening chamber door and removing sterilized items. The pressure drops very quickly to 0, but takes a while for the temp. to drop.
14. Take care when opening chamber as hot H2O will drip from the underside of the door when opened. Make sure that H2Odoes not splash on the control panel.
Hirayama HV-110 (NSC 528) large blue autoclave
1. Warning: this autoclave does not take as long to heat up as the above Sanyo, but does take a little while.
2. Use distilled H2O in autoclave.
3. Check to see if there is any H2O in the main chamber. To do this, wiggle the whole unit and look down in the chamber – if you see H2O through the round hole, do not add anymore H2O. There should not be any standing H2O in the bottom of the chamber.
4. Remove the exhaust tank and fill with H2O to the High level mark.
5. Open the chamber door and pour H2O inside (~2 L) until you can physically see H2O in the round hole in the bottom of the chamber. This will take the H2O level in the exhaust tank to usually between the HIGH and LOW mark.
6. Fill the exhaust tank with H2O to the LOW mark and not over the HIGH mark. Connect the exhaust hose and place the tank inside the unit.
7. The control panel is usually in “Sleep” mode – blank panel. Hit any button to take it out of Sleep mode.
8. Turn the power switch on, lower right in body of sterilizer.
9. Place the items to be sterilized into the chamber. Push the lever on the front of the door to locked (be sure to push the lever all the way or the control panel won’t work). You can hear it engage.
10. Mode. Mode (keep pressing Mode to select) – select either liq. Solid or agar (please note that the temp. for agar is 60 oC, this is hot enough to melt agar but does NOT sterilize).
11. Func – select either timer, clock. Print is one of the choices, but there isn’t a printer.
12. Func./Timer/Set/Next/use arrow to set hr. if that’s how you want to time/or next/use arrow for min./Enter
13. Settings. Temp. - usually 121 oC; or time - there is a lag time for the sterilizer to reach the set temp. and this depends on the volume you are autoclaving. For example, for a 500 ml flask, there is a 15 min. lag, so for a 15 min. sterilization ,set the time for 30 min.).
14. Press the start button to begin automatic sterilization process.
15. Take care when opening chamber as hot H2O will drip from the underside of the door when opened. Make sure that H2Odoes not splash on the control panel.
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This information is given as a guide to the facilities and instrumentation available in the DNA/Protein Core facility at Georgia State University. If you have any concerns or thoughts about the content of this website please contact: John Houghton (404) 413-5390
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