How to Plate Fungal Samples Frozen in Glycerol

Step-by-Step Guide

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   Step 1: Put on your gloves!

   This includes lighting your burner and checking to see that the blower is on for the lab bench. ,,,,,,, Dip the loop into the 95% ethanol and flame it. ,,, Quickly stick it into the frozen glycerol sample.
   
   This may take several attempts to fully get the end of the loop into the sample. , This will most likely take 2 or more dips into the sample. , Close the Petri dish and the sample. , Make sure the ice is on the samples and close the Styrofoam container. ,,
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   Step 2: Layer the bottom of the Styrofoam container with ice packs from the fridge

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   Step 3: 1 layer is fine.

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   Step 4: Retrieve the box of glycerol samples and place it in the foam container.

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   Step 5: Cover the box with 1 or 2 more ice packs and close the container.

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   Step 6: Set up your lab bench.

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   Step 7: Spray down the lab bench with 70% ethanol and wipe it with a Kimwipe.

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   Step 8: Spray gloves with 70% ethanol and lather until dry.

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   Step 9: Place the foam container next to your feet.

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    Step 10: Open the foam container and the sample box and pull out the first sample

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    Step 11: immediately put the extra ice packs on top of the rest of the samples and close the Styrofoam container.

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    Step 12: Label the first Petri dish accordingly.

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    Step 13: Open the sample and place the cap upside down on the bench.

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    Step 14: Hold the sample in your left hand and with your right hand.

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    Step 15: Start at the end closest to the handle and flame the loop until it’s red; work your way down to the end of the loop.

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    Step 16: Open the Petri dish with your let hand while still holding onto the sample.

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    Step 17: Make sure the tip of the loop is very red.

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    Step 18: When you can clearly see sample on the loop

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    Step 19: lightly spread it first on the filter paper

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    Step 20: and make sure it’s fully covered

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    Step 21: then continue to lightly spread it on the rest of the Petri dish (You do not want to scrape the agar).

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    Step 22: When the entire Petri dish is covered

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    Step 23: put the loop into the 95% ethanol container.

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    Step 24: Place the sample back into the box.

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    Step 25: Parafilm the Petri dish and set it aside.

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    Step 26: Repeat steps 8 -17 as many times as necessary.

Detailed Guide

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