Alban Tomaj ; Meaghan Murphy ; Kylie Yoshimura

Began: 07 April 2011 ; Completed: 11 April 2011

Genetics Lab – Doe
Phage Assay

Purpose:
The purpose of this laboratory experiment is to measure the amount of plaques produced by T4
bacteriophage (virus) on bacteria, particularly E.coli. Bacteriophage infect bacteria by transferring it's
DNA into the bacteria's DNA. The infected bacteria then translates the new viral DNA, creating viral
components for the construction of viruses. After a number of viruses are constructed, the cell lyses,
breaking apart and releasing the new phage into the medium, forming a plaque on the agarose gel. By
using different concentrations of the T4 phage, we can perhaps create a correlation with the amount
phage and the amount of plaques produced. We will conduct this experiment by creating different
dilutions of the phage with saline solution. These different dilutions include 10-5 ,10-6 , 10-7, and 10-8.
The experiment was conducted using sterile techniques in order to avoid contamination. The E.coli
used was in a 10-1 concentration.

Results:
After at least 24 hours, our group met and counted the number of plaques produced from the
lysis of the cells. Table 1.1 provides a summary of our results. In the 10-5 plate, we counted 73 total
plaques; however, due to human error, the dish was not completely covered with the phage and E.coli.
As a group we assumed a total of 140 plaques if the plate was completely covered (this is the only
plate where we experienced this problem). In the plate with 10-6 dilution, we counted 44 plaques. This
was lower than our first dilution which is what we had expected. We noticed, that these plaques were
larger than the rest of the plaques in the other plates. We counted 64 plaques in the 10-7 dilution
dilution. The plaques appear smaller. Lastly, the last plate which had the 10-8 dilution had a total of 89
plaques. Again, the plaques appear smaller.
We experienced an inverse correlation with the titer; as the dilutions increase (concentration
becomes smaller), the titer increases.

Table 1.1 – Result of Phage Assay Experiment

Dilution Total Plaque(s) Titer (#plaques/dilution factor)
10-5 73 7.3 x 105 730,000
-6 6
10 44 4.4 x 10 4,400,000
-7 7
10 64 6.4 x 10 64,000,000
10-8 89 8.9 x 108 890,000,000