Smolke:Washroom Procedures

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Sterilizing bacteriological items

Sterilizing Bacteriological Plates

Wear gloves. Remove bag from bin. If necessary, make a second double bag and add plates to that bag so it isn't overfull for the autoclave process. Secure with autoclave tape, but leave a gap so air can enter the bag during the autoclave procedure.

Double bag two bags and put into white bin by entrance to room B17.

Place one to two bags in autoclave tray on lower rack. Remove extra rack. Fill the tray with about 1 to 1.5 in of water. IMPORTANT: Ensure the closures are upright ! As the autoclave process proceeds, the bag will collapse and liquid contents may escape into the autoclave resulting in a big mess.

Run the bags on the liquid 30 min cycle. Dispose of bags in regular waste containers.

Check the contents of the container each day, but you should only have to run the cycle M,W,F to stay on top of it.

Sterilizing Bacteriological Cultures in Test Tubes

Wear gloves. Remove caps from tubes and for a full rack, invert the silver basket over the open tubes. Holding firmly, invert the contents into the sink with running water. Leave the white colander in the sink to catch tips that may come out.

Invert again and remove basket. Hold the rack under running water to fill tubes. Drain, repeat once more, but at end of drain, leave tubes inverted in basket and remove tube rack. Check the bottom of tubes for any remaining debris, and if necessary, rinse once more before loading into the wash basket.

Secure tubes in basket with movable panel and run through the washer.

Rinse the tube rack and leave to dry during the wash.

Wash caps in large bucket in sink by soaking with a small amount of washer detergent. Pour into rinse basket and rinse well. Shake excess water in sink and air dry. You can speed this process with leaving in the warm autoclave if you like.

With clean hands, inspect the tubes when wash is finished for any remaining debris. You can leave briefly in the autoclave to help dry the tubes if you like. Careful if you do this, as the whole rack will get hot!

Load tube rack and cap each tube. Add autoclave tape across caps on full racks.

Run on Tips cycle. Store on shelf above ice machine.

Sterilizing Bacteriological Glass Plating Beads

Wear gloves. Collect glass beads from the benches in a 2 liter glass beaker. Gently tap the plastic containers to loosen beads if necessary. Note the volume of beads to wash. Each bottle will hold about 60ml of beads, so estimate how many bottles you will need now.

In the washroom, add hot water and two squirts of pink hand soap into the glass beaker. Agitate with a spatula from the weigh room. Soak for at least 15 min.

Wear gloves and use a sink in the Lab. Pour the beads into the sieve draining the soapy water. As they drain, rinse the glass beaker with distilled water to remove remaining soapy water. Rinse the beads well with distilled water. Agitate the beads with your gloved hand, shake and let drain.

With the glass beaker in the sink, use your two hands to gently guide the beads from the sieve back into the glass beaker. Pick up any errant beads from the sink and rinse well.

In the chemical hood, pour to cover the beads, with ethanol. Return the container to storage after finishing. Carefully travel back to the washroom with the beads in the ethanol. Let sit for a few min and agitate briefly with the spatula.

Gently drain the beads through the sieve into the sink. Shake the sieve to remove excess ethanol. Briefly leave the sieve to drain on the counter on some paper towels. Then run some water to rinse the excess ethanol down the drain. You can also leave the beads to dry briefly in the autoclave. Do this for a few min. with the door open to help evaporate the ethanol.

While the beads are draining, assemble the appropriate number of 100ml glass bottles, one large funnel, and a small glass beaker. With the small glass beaker, scoop beads from the sieve and into a glass bottle using the funnel, filling each bottle to the 60ml mark. When there are too few beads to scoop, gently pour the remaining beads utilizing the funnel.

Leave beads overnight on the counter with the caps partially on to facilitate complete drying.

Leaving caps loosely on the bottles, tape with autoclave tape and run on the next tips cycle. When cool, completely sure caps and store in the drawer by B17.

You will usually have to keep up with this about once a week, taking into consideration the overnight drying time.

Sterilizing Toothpicks for picking Cultures

Fill toothpicks in small glass beakers. Make a foil cap by folding over aluminum foil two to three times for durability.

Add autoclave tape to cap and run on tips cycle.

When cool, add to drawer by B17.

I check this stock when doing the beads.