Autoclaving tips for success

• Use an autoclave safe tray or bin to put your materials in when autoclaving them. If agar boils over, the liquid will end up in the tray, not on the bottom of the autoclave (where it will clog up the drains).  Trays or bins are also good for supporting autoclave bags. Please make sure the tray or bin is autoclave safe and can handle high temperatures, otherwise it may melt all over the autoclave.

• Autoclave bags:  prior to autoclaving, add 100-200 mL water to the bag and then loosely close it.  This will help generate steam inside the bag, which will greatly aid in destroying microorganisms.

• Don’t fill bottles or tubes more than 2/3 full to prevent boil-overs.

• Keep lids and caps loosely closed.  Tightly sealed bottles may explode.

• Use only Type I borosilicate glass (Pyrex) or autoclavable plastic containers.  Do not use old food jars (peanut butter, pickles…) to autoclave materials because they will break under autoclave conditions.

• Autoclave your materials for the correct time period (see Autoclaving Fact Sheet for more information). 

• Be sure to close the door tightly.  If the door is not closed tightly, steam will leak from around the door and the autoclave will make screaming noises.  If this happens, abort the cycle (reset the autoclave), tighten the door, and try again.

• When you start a cycle, stay and watch for a few moments to make sure the autoclave begins the sterilization cycle.  If there is a problem with the steam, you will know at this point, instead of coming back an hour later to find that your items are not sterilized because the autoclave didn't’t run through the cycle.

• Do not open the door until the pressure is down to 0 psi.  When you open the door, only open it about one inch at first—this helps ensure that you won’t get blasted with steam.

• Do not stand in front of the door when you open it.  Stand behind the door to avoid getting blasted with steam.

• When loading and unloading the autoclave, always wear heavy duty gloves to protect your hands from getting burned.  Remember, the inside surfaces of the autoclave are hot, even if you haven’t run a cycle yet. 

• Never wear shorts, short skirts, open-toed shoes or sandals when accessing items from the autoclave.  If items boil over or shatter, your legs and feet may be severely burned.  Wear a rubber or waterproof apron and face shield if possible.

• The door of the autoclave was not meant to hang open (it is hard on the hinges).  So, when you are finished using the autoclave, please reseat the door.

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