Cleaning and autoclaving guidelines

Washing glassware and tools

We dishwash all glassware using the dishwasher in the autoclave room (56-414). When using the dishwasher, fill out your information (date, time, name, phone number, office number, and PI initials) in the dishwasher log on the metal racks to the left of the autoclave. The standard cycle is sufficient, and it takes 90 minutes to run.

For any items that need to be washed by hand, we use Alconox as our soap. Alconox is stored as a powder underneath the sink. Dilute this powder in water in the squirt bottle labeled “Alconox”. There is no precise measurement for this.

Glass bottles and flasks

Note

If you empty a glass bottle or flask (media, PBS, gelatin, ELGA water, etc.), you are responsible for bleaching and rinsing it. If you cannot bleach and rinse the bottle immediately, keep the bottle on your personal lab bench.

  1. If the glassware had cells in it (e.g., midiprep culture) first add bleach to a concentration of 10% and let sit for at least 15 min.

  2. Rinse the glassware to remove the bleach.

  3. Glassware can now be placed in the area to be dishwashed, to the left of the Elga water machine. Alternatively, follow the steps below to wash by hand.

    1. Add Alconox and water, and shake/swirl vigorously to clean. Use the brushes by the sink for tough spots.

    2. Rinse thoroughly with tap water until no soap bubbles are left.

    3. Perform a quick rinse with DI water.

    4. Put glass to dry on drying rack.

  4. Place clean, dry glassware from the dishwasher or drying rack on the shelf labeled 'Clean glassware' outside of the TC room. To prevent dust accumulation, store beakers upside down and add foil to flask mouths.

  5. For sterility, flasks that will be used to grow cultures are additionally autoclaved. Place autoclave tape on the foil on the flask mouth, then run the flasks in a typical dry autoclave cycle (see below).

Glass beads

Note

The dirty bead box should always be filled with 10% bleach

  1. If bleach levels are low, add more bleach to dirty bead box and allow to sit for at least 15 minutes. Dump out beads and bleach into mesh strainer found in cabinet to the left of the sink.

  2. Soap with Alconox and then rinse with tap water.

  3. Perform a quick rinse with DI water.

  4. Let beads fully dry in strainer overnight.

  5. Use funnel to put beads into glass bottle for autoclaving.

Dissection Tools

Note

Dissection tools should be washed as soon as dissection is finished. Dirty tools should not be left at autoclave station.

  1. After dissection, spray with ethanol and remove any visible residue.

  2. Wash with Alconox and then rinse with tap water.

  3. Perform a quick rinse with DI water.

  4. Let dry and wrap in tin foil.

  5. Place back into metal container and move to area next to ELGA water dispenser to be autoclaved.

Autoclaving

Glassware and plastics are autoclaved as needed. Usually, this amounts to twice a week. Clean glassware on the appropriate storage shelf (i.e., wooden shelves next to the entrance to main TC) will be autoclaved as well as any empty tip or tube boxes in the autoclave area next to the sink. Prep materials as described below and place in an autoclave-safe bin (located on the rack to the left of the sink). The autoclave is located in room 56-414 and requires card access.

Important

Do not leave heat-proof gloves in the autoclave room. These tend to disappear… However, it is okay to leave the cart in the room.

Autoclave Operation

  1. Ensure there is not a program currently running. If someone else is currently running the autoclave, fill out your information in the autoclave log (date, name, phone number, office number, PI initials, and cycle time) on the metal shelves and write “next” to the left. Once the autoclave is free, cross out “next” and fill in the start time for your desired cycle.

Note

Only write “next” if no one else has done so already. This keeps the queue at one person. If someone has already written “next”, come back later.

  1. Open the door by turning the wheel fully counterclockwise, then turn the lever counterclockwise.

  2. Place the bins on the racks inside the autoclave and close the door, first turning the lever and then the wheel fully clockwise.

  3. Using the screen on the autoclave, ensure the jacket is turned on and select your desired cycle:

    • For dry cycles (glassware, plastics, etc.): Gravity >> 20 min / 20 min (runtime ~45 min)

    • For wet cycles (liquids such as PBS, LB, etc.): Liquid >> 30 min / 0 min (runtime ~1 hr)

  4. When the cycle is complete, open the door by first turning the wheel counterclockwise and then the lever. Be cautious of hot steam escaping from the open door.

  5. Remove bins from the autoclave using heat-proof gloves.

  6. Close the autoclave door using the lever, but do not tighten the wheel when the autoclave is not in use.

Tip boxes

  1. When a box of tips is emptied, recycle the empty plastic insert and throw away any autoclave tape, then place the empty box in the area next to the ELGA water dispenser.

Note

Filter tip boxes do not need to be reused and the whole thing can be recycled when empty. Recycle any broken tip boxes.

  1. Add inserts with new tips to empty boxes.

  2. Add strip of autoclave tape where the lid meets the side of the box.

Tube containers

  1. When a container of 1.7 or 0.6 mL tubes is emptied, throw away the autoclave tape and place the empty container in the area next to the ELGA water dispenser.

  2. Fill with new tubes.

  3. Screw on top of container loosely to avoid warping while in the autoclave.

  4. Add strip of autoclave tape where the lid meets the side of the container.

Glass pipette containers

  1. Make sure no glass shards remain in metal container (shake out over sharps container), throw away the autoclave tape, and place the empty container in the area next to the ELGA water dispenser.

  2. Carefully fill metal containers with glass pipettes with the thin end pointing down (i.e. larger handle side points outwards when you open it).

  3. Add strip of autoclave tape where the lid meets the side of the container.

Toothpicks

  1. When a container of toothpicks is emptied, throw away tin foil and autoclave tape and place the empty container in the area next to the ELGA water dispenser.

  2. Fill container with toothpicks.

  3. Cover with foil and mark with autoclave tape.

Flasks

  1. Add strip of autoclave tape to the top of clean flasks covered with tin foil.

Dissection tools

  1. Make sure tools are clean, dry, and individually wrapped in tin foil inside their metal container.

  2. Wrap the outside of the metal container with tin foil and mark with autoclave tape.

Glass bottles (containing liquid or glass beads)

  1. Loosely screw on bottle cap.

Important

Do NOT screw caps on all the way!

  1. Use autoclave tape to further attach the cap to the bottle.

  2. Cover cap and top threads with tin foil. This helps keep the edges of the top sterile while the cap is not fully screwed on.

  3. Mark the top of the cap with autoclave tape.