Autoclaving

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Autoclaving

• By Laurie Brier

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The Autoclave
Pressure gauge
Light
Timer
Temperature Gauge
Exhaust Selector
Door
Latch
Handle
Drain Switch
http//biology.clc.uc.edu/fankhauser/Labs/Microbio
logy/Media_Prep/Autoclave.htm
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Materials Needed

• Materials to be autoclaved
• Autoclave tape
• Autoclave bucket

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Preparing the Autoclave Destined Materials

• Twist any bags of materials shut, loosely
  wrapping the autoclave tape around the neck of
  the twist.
• Loosen the caps of all bottles, and place a piece
  of tape onto the cap of each bottle.
• Place materials into the autoclave bucket
• DANGER if anything is closed too tightly in the
  autoclave it will burst!

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Autoclaving Step 1

• Make sure that the drain is closed
• Fill the autoclave with distilled water
• DO NOT fill past the mark labeled inside of the
  machine

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Drain Setting

• Found underneath autoclave door latch.

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Step 2

• Place the bucket of prepared materials into the
  autoclave

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Step 3

• Shut and latch the autoclave door

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Step 4

• Located right above the door.

• Set the exhaust selector to fast or slow.
• Setting will depend on the type of materials you
  are autoclaving.

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Step 5

• Set the autoclave for 25 minutes (or longer
  depending on the volume of liquids).
• The pressure and the temperature inside of the
  autoclave will begin to rise
• This can be observed from the gauges located at
  the top of the autoclave

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Temperature and Pressure Gauges

• Top left of autoclave

• Top right of autoclave

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Step 6

• After the set time has passed and the pressure
  gauge falls to zero, unlatch and open the
  autoclave.
• Steam will billow out of the autoclave, once
  opened.
• Wear gloves and long sleeves for protection.
• Remember to stand back while opening.
• Warning
• DO NOT attempt to open the autoclave before the
  pressure has dropped to zero as serious burns
  could occur.

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Step 7

• Using gloves, remove the bucket from the
  autoclave.

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Step 8

• Check the autoclave tape for darkened areas
• You should either see stripes or the word
  sterile on the tape
• If the tape did not darken, your items are not
  sterile
• Additional time in the autoclave may be needed