According to a protocol developed at John A. Burns School of Medicine, JABSOM, Hawaii, the first step during an autoclave procedure is choosing one of the three available programmed cycles. Cycle one lasts 25 minutes and is used for sterilizing dry articles such as clean glassware; cycle nine lasts two hours and is used to decontaminate materials; cycle 10 lasts for 15 minutes and is used to sterilize liquids. Technicians should be careful to place the material to be sterilized into the cart, never directly onto the chamber floor.
This protocol is used to sterilize liquids at 249.8 degrees Fahrenheit for 30 minutes. First, technicians must perform the machine setup, filling the autoclave water reservoir with distilled water to cover the heating coils. Then, they should load the samples into a secondary container, directly onto the metal grate. They should then set the temperature knob two notches above the 121 Celsius, or 250 Fahrenheit, mark and set timer to 55 minutes. Technicians should check the temperature gauge -- 25 minutes into the run -- to make sure it has reached 250 degrees F. When the operation finishes, wait 25 to 30 minutes before opening the door.
According to the Department of Chemical & Biomolecular Engineering at the University of Maryland, during an autoclave protocol, technicians should observe some important procedures including: checking if the door is properly closed to avoid steam leaking; opening the condensate valve and the steam valve supply; placing items in the chamber; locking the autoclave door; introducing steam into the chamber; autoclaving for 20 to 30 minutes, and terminating the cycle by shutting off steam to the chamber; unlocking and opening the autoclave door when the needle of the chamber pressure indicator is in the area marked around zero; removing autoclaved items from the chamber and cleaning up.
According to this protocol, technicians should first close the drain valve and pour about one gallon of hot water into the reservoir at the bottom of the autoclave. Then, load the material to be autoclaved, close the door and latch it by pressing down firmly. Next, they should select the appropriate exhaust or cycle -- slow for water-containing materials, fast for laboratory apparatus. After selecting the cycle, wait until it is finished then open the door only when pressure has returned to zero. Finally, remove the autoclaved items with the aid of gloves and open the drain valve to empty the water reservoir.