Autoclave Procedure - Cuddington-Lab/documents GitHub Wiki

Every week the jars and beakers used for the Duckweed Experiment need to be sterilized via autoclave.

This procedure can only be performed by graduate students, or by undergraduate students while supervised by graduate students

To prepare for autoclave:

Scheduling the autoclave

  1. Go to the booking website

Big autoclave in B2-354: https://uwaterloo.ca/biology/services-and-resources/molecular-biology-core-facility/molecular-biology-core-facility-mbcf-equipment-booking

Small autoclave in B2-354: https://outlook.office365.com/owa/calendar/[email protected]/bookings/

  1. Select the relevant autoclave
  2. Enter your uWaterloo user name
  3. Select "Gravity 2: Sterilize 45min; dry 20 min; 1 hour 30 minutes"
  4. Select date and time

Running the autoclave

  1. Prepare items for autoclave. Jars should be loosely capped, with a strip of autoclave tape applied on the cap. Beakers should be loosely covered with aluminum foil, with a strip of autoclave tape applied to the foil. Metal utensils such as tweezers and scoops can be wrapped in aluminum foil, again with a strip of autoclave tape. Write "B2-241" on each strip of tape.
  2. Place everything in the white plastic tray. DO NOT use any other trays.
  3. Bring the tray to the autoclave room, B2-354. Use the cart and take the elevator in the STC connection, DO NOT carry it by hand. Going up and down stairs with a tray full of glassware is an accident waiting to happen.
  4. Fill in the sign in sheet with your name, start time, selected cycle (Gravity), length of cycle, supervisor (Cuddington) and extension number (33669).
  5. Unlock autoclave door by turning rear lever to the left and pull open door cautiously. Autoclave should be empty. Place tray inside autoclave, ensure no other items are in the tray, except those being autoclaved.
  6. Before closing, make sure all jars and beakers are on their side and slightly open. This ensures air can enter and circulate. The autoclave works by circulating high-temperature steam to sterilize surfaces—if the steam can't reach the inner surfaces of a beaker, it can't sterilize it! (The utensils are okay since they are thin, small, and metal; the heat can easily penetrate the wrapping and sterilize them.)
  7. Close autoclave door and turn rear lever to the right to lock the door. Rotate front wheel clockwise till tightened (as much as you can personally tighten it).
  8. Touch the autoclave screen and select Gravity >> Gravity 2 >> confirm door is closed >> start. Wait for the chamber temperature to reach the target temp and make sure steam is not leaking out of the door. If it is, you need to tighten the wheel more. Once you are sure steam is not leaking and the chamber has heated up, you can leave and wait for it to finish. Gravity 2 takes 65 min total for the sterilizing and drying process.
  9. Return to autoclave room after 65min (don't forget the cart); when the screen reads "CYCLE COMPLETE" and timer is up, wait 3-4 additional minutes for jacket temperature to lower.
  10. Rotate wheel in counter-clockwise direction till loose; rotate the rear lever to the left to unlock the door and pull open cautiously, there will be heat and steam leaving the autoclave as this is done - try to stand clear of the direct opening of the door
  11. Wear autoclave gloves and remove tray from autoclave cautiously - the tray and inside of the autoclave will still be hot
  12. Close the autoclave door and rotate the rear lever to the right to lock
  13. You may now place jars and beakers upright so they do not roll around later. You can also tighten the foil on beakers, but be careful since it's still hot.
  14. Bring tray back to lab once it is cool enough to handle. Don't remove the jar caps or tin foil off the beakers (this ensures the inside remains sterile). Note: the autoclave tape will have black stripes, indicating the marked material has been autoclaved

Note if autoclaving in B1-376: empty the condensation bottle (which is on the left side of the autoclave) after you finished the autoclave in B1-376 before you leave, otherwise the water will overflow.

Contact person for questions: Akash Jairaj, MBCF co-ordinator ([email protected])