Documentation:Standard Operating Procedures for Tuttnauer Autoclave 3870

Standard Operating Procedures for Tuttnauer Autoclave 3870

1. Please obtain proper training from the Faculty technician prior to using the autoclave.

2. Sample/waste handling prior to autoclave:

Biohazardous wastes should be placed in clear autoclave bag, with an autoclave indicator tape placed in a visible location. The bag should be loosely open to allow steams in and be placed in a metal tray.

Samples should also be placed in metal tray in case of spills.

3. Before operating the autoclave, check the water level from the top reservoir. It is important to have enough water but at the same time the water does

not cover the bar where steam is produced. The water level should reach the metal ring but do not cover the bar. Distilled water should be used to fill

the reservoir.

4. Place a metal tray in the chamber and load your samples in the metal tray.

Note: It is important that you always use a metal tray to contain your samples in case of any spillage.

5. Turn the control knob (bottom of the panel) from 0 to “fill water”.

You will observe water starting to fill the bottom of the chamber.

6. When the water reaches the line at the front of the chamber, turn the control knob from “fill water” to “Sterilize”. The water should stop.

7. Now you can close the door and shut tight. Turn clockwise to close and turn counterclockwise to open.

8. Set the timer to 60 min, keep the temperature at 121^oC and then turn ON the autoclave.

9. Wait for approximately 25 - 35 min until the pressure gauge reads above 15 psi. Both the red and white needles will go up to reach maximum

pressure.

10. When the pressure of over 15 psi is reached, turn the timer to 15 min for sample sterilization (60 min for waste treatment).

11. For each autoclave run, you must sign the logbook.

Note: You must sign one line per run.

12. When the sterilization time is up (timer back to 0), it automatically proceeds to depressurize slowly.

Biohazardous wastes - turn the control knob to “ExH/DRY” to quickly depressurize

Equipment and glassware - turn the control knob to “ExH/DRY” to quickly depressurize

Samples/media – leave unchanged until pressure gauge reads 0 psi.

Note: It is important to let the pressure slowly releases for any liquid containing sample preparation as the “ExH/DRY” function will drop

the pressure too quickly and lead to spillage from the flasks/bottles.

13. Wait until the chamber pressure drops to 0 psi. The white needle will continue to drop down to 0 psi while the red needle will remain at the highest

reached pressure.

14. Turn the control knob to “0”.

15. Open the door to release remaining pressure.

Note: You should stand clear of the autoclave door as steam will be burning hot!

16. With an oven mitten, unload your samples from the autoclave.

17. If you have further samples/wastes, turn the red needle on the pressure gauge back to 0 psi, then return to step 3 and repeat.

18. If you are done with the autoclave, make sure to turn the red needle back to 0 psi, then turn OFF the autoclave.

Note: It is important to turn the red needle back to 0 psi so it rises to the maximum pressure for next run.

19. At this point, please make sure you have signed the log book or you may not be allowed to utilize the autoclave in the future. Report any anomalies

of the autoclave to the Faculty Technician.