Serial Dilution

Introduction: The Purpose of serial dilution is to determine the number of bacteria per unit volume in the original culture, determination of the culture density in cells per ml. Once the culture has been diluted it can be spread on agar plates. Agar plates allow for individual bacterial cells to be separated spatially. If done correctly, there is a low probability of having two cells very close to each other. When each of these spatially separated cells multiplies, spatially separated colonies are formed. We call these clonal colonies as each visible colony arose from a single progenitor cell. The number of colonies observed is thus a direct measure of the number of bacteria spread on the surface of the plate.

Serial Dilution: Gathering the tools

1. Stock Culture 2. Dilution tubes or jars (make sure to label them) 3. Pipetter 4. Agar Plates 5. Spreader

Creating a Serial Dilution

1. 2. 3. 4. 5. Place 9ml of media in each dilution tube or jar Use the Pipetter to add 1ml of stock culture to the rst dilution tube. From the rst dilution tube use the Pipetter to add 1ml from the rst tube to the second tube. From the second tube use the Pipetter to add 1ml from the second tube to the third tube. Continue until you reach the desired dilution.

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Plating the Diluted Bacteria

1. Use the Pipetter to place a sample of the diluted culture on the Agar plate. 2. Push a spreader across plate to distribute the sample
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