Cleaning and autoclaving guidelines

Washing glassware and tools

We dishwash all glassware using the dishwasher in the autoclave room (56-414), or wash item by hand using Alconox. Both are described below. The cleaning procedure for glassware, glass beads, and dissection tools are also described below.

Operating the dishwasher

  1. Check that the dishwasher is free and fill out the log (date, time, name, phone number, office number, and PI initials) on the metal racks to the left of the autoclave.

  2. Confirm that there are enough detergents in the large containers on the floor to the right of the dishwasher.

  3. Prepare glassware as described below.

  4. Load glassware into the dishwasher.

  • If needed, the shelves/racks in the dishwasher can be swapped.

  • Avoid overfilling the dishwasher, as this can lead to glassware hitting each other during the run and breaking.

  • Place bottle lids in the metal mesh basket, and fit the mesh lids snugly over top to secure them in place. Otherwise, the lids will fall to the bottom during the cycle.

  1. Select the “B2 Standard” cycle and press the button (play button icon) to start the cycle. The cycle takes 93 minutes to run.

  2. When the cycle is complete, remove the glassware from the dishwasher. Note that it will be hot!

  3. Store clean, dry glassware in lab as described below.

Washing glassware by hand

For any items that need to be washed by hand, we use Alconox as our soap.

  1. As needed, dilute Alconox powder in water in the labeled squirt bottle by the sink (next to the Elga water dispenser). Alconox powder is stored under the sink, and there is no precise measurement for mixing it with water (a little bit of powder is sufficient).

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

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

  4. Perform a quick rinse with Elga water.

  5. Place glassware on the drying rack above the sink to dry.

  6. Once dry, remove promptly to free up space and to reduce the chance that it will be accidentally knocked off the drying rack.

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. Remove the tape label from the glassware. A razor blade can help to remove stubborn tape fragments—if these are left on when dishwashing, they become even harder to remove!

  4. Glassware can now be placed in the area to be dishwashed, to the left of the Elga water dispenser. Alternatively, follow the steps above to wash by hand with Alconox.

  5. 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.

  6. 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). Running the dry autoclave cycle is typically a separate lab job from dishwashing.

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 Elga water.

  4. Let beads fully dry in strainer overnight.

  5. Use funnel to put beads into glass bottle for autoclaving on a dry cycle.

Dissection Tools

Note

Dissection tools should be washed as soon as dissection is finished. Dirty tools should not be left at the 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 Elga water.

  4. Let dry and wrap tools individually in tin foil.

  5. Place tools back into the metal container and wrap the whole container in foil. Add autoclave tape.

  6. Place the container in the autoclave area next to the Elga water dispenser, to be run in a dry autoclave cycle.

Autoclaving

Glassware and plastics are autoclaved as needed, usually twice a week as part of a lab job. 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 Elga water dispenser. Prep materials as described below and place in an autoclave-safe bin (located on the rack to the right of the sink). The autoclave is located in room 56-414 and requires card access and completion of a training module.

Operating the autoclave

Note

When running the autoclave, it is okay to keep a cart in the room, pushed out of the way, since we have two carts. However, do not leave heat-proof gloves in the autoclave room, otherwise they might disappear.

  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 (starred cycle, runtime ~45 min)

    • For wet cycles (liquids such as PBS, LB, etc.): Liquid >> 20 min / 0 min (starred cycle, runtime ~45 min)

  4. When the cycle is complete, remove items promptly to be courteous to other users. 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.

  3. Autoclave on a dry cycle.

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 a strip of autoclave tape where the lid meets the side of the container.

  5. Autoclave on a dry cycle.

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.

  4. Autoclave on a dry cycle.

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.

  4. Autoclave on a dry cycle.

Flasks

  1. See instructions above for washing glass flasks. Clean, dry flasks should be stored covered with foil in the “Clean glassware” shelf next to TC.

  2. Add strip of autoclave tape to the foil on the top of the flasks.

  3. Autoclave on a dry cycle.

Dissection tools

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

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

  3. Autoclave on a dry cycle.

Glass bottles containing liquids or glass beads

  1. Prepare solutions as described in the recipe protocol, or glass beads as described above.

  2. Loosely screw on bottle cap.

Warning

Do NOT screw caps on all the way! This could cause an explosion in the autoclave.

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

  2. Cover cap and top threads with foil. This helps keep the edges of the top sterile while the cap is not fully screwed on, so make sure the foil is snug around the threads.

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

  4. Autoclave bottles containing liquids on a liquid cycle, or glass beads on a dry cycle.