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Serial Dilution Method for Bacterial Counting

1. The document describes a procedure to serially dilute a bacterial sample to allow for easier counting of bacterial colonies when plated. 2. Bacteria was collected from a person's hand via swab and placed in the first of 7 test tubes containing water. Serial dilutions were performed by transferring 1 mL between tubes. 3. Samples from each tube were plated and incubated, then colonies were counted. Fewer colonies were observed in more diluted plates, showing the dilution method was effective.

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3 views2 pages

Serial Dilution Method for Bacterial Counting

1. The document describes a procedure to serially dilute a bacterial sample to allow for easier counting of bacterial colonies when plated. 2. Bacteria was collected from a person's hand via swab and placed in the first of 7 test tubes containing water. Serial dilutions were performed by transferring 1 mL between tubes. 3. Samples from each tube were plated and incubated, then colonies were counted. Fewer colonies were observed in more diluted plates, showing the dilution method was effective.

Uploaded by

Holian
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOC, PDF, TXT or read online on Scribd

Title

Objective

To decrease a bacterial concentration to a required concentration in able to count easily and


accurately when the bacteria is plated to an agar plate.

Materials

Test tube with sterile water

Petri dish with agar

Pipette dropper

Alcohol lamp

Sterile swab stick

L-shape spreader

Tape

Plastic wrap

Procedure

1. Prepare all the materials needed for isolation of the bacteria.

2. Prepare 7 test tubes. Put 10 mL sterile water in the first test tube and 9 mL on the
following six test tubes. Label the test tubes from 1 to 7.

3. Using a sterile swab stick, swab the hand of the person who travelled from Batangas to
Indang. Make sure to swab even between the fingers.

4. Put the sterile swab stick in the tube 1. Observe and apply aseptic technique.

5. Using the pipette dropper, transfer 1 mL from tube 1 to tube 2. Now, the tube 2 has 10 mL
while the other tube remains 9 mL. Again, observe and apply aseptic technique.

6. Repeat the procedure in transfering from one tube to another until the tube 7 has 10 mL.

7. Get 0.5 mL from tube 1 using pipette dropper and place it inside the petri dish with agar.
Observe aseptic technique. Spread it gently with the use of L-shape spreader.

8. Repeat the procedure to other tubes. Do not forget to apply aseptic technique.

9. Label the petri dish from 1 to 7. Sealed the petri dish with tape and plastic wrap.
10. Incubate at room temperature for 24 hours.

11. Observe and count the colonies present.

12. After counting the colonies, place it inside the refrigerator to stop growing and
multiplying.

Result

Plate Number No. Of Colonies Used to spread the


(Bacteria) bacterial culture

1 8 Sterile swab

2 14 L-shaped spreader

3 L-shaped spreader

4 10 L-shaped spreader

5 20 L-shaped spreader

6 1 L-shaped spreader

7 0 L-shaped spreader

Conclusion

Following the proper procedure or aseptic technique when isolating bacteria is very essential
in order to have an alive bacterial colonies, a better view for lab results and a successful
experiment.

As being assigned to culture the first sample, it is expected for our plate to have a more
numbers of colonies than the following plate. But, some plates have a higher number of
colonies compared from our plate. Based from pur perspective, it may be caused by
improper aseptic technique, moist inside the plate that may be killed some of the bacteria,
or may be improper storage. But the more clear caused is method in spreading the bacteria.
The more set of streaks we perform, the more isolated colonies we get. We observed how
serial dilution affects colonies of bacteria having given rise to fewer colonies. So, we
obtained 8 for the plate 1,14 for the plate 2, 3 for the plate 3, for the plate 4, 20 for the plate
5, a big clamp of colony for the plate 6, and 0 for the plate 7. The reason why we did not get
the expected higher number colony for Plate 1 is because of the streaking. However, the
overall results confirmed that the dilution factor was preserved till the last dilution. To
improve “serial dilution experiment” it is important to use several plates (2-3) for each
diluted tube, so we could obtain the average from these plates that could lead to more
accurate results in calculations of the concentration of bacteria in the original culture.

Reason

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