Research Question:

To find out how the presence of bacteriophages in the soil around red or white oak trees has a correlation with the health condition of oak trees.

Rationale:

A Plaque Assay helps us determine if there is a presence of bacteriophages by adding Arthrobacter directly to the lysate. We can tell the existence of bacteriophages by checking the presence of plaques on the agar plate. Samples were collected to expand the data size and provide more insight into whether the presence of Arthrobacter phages in the soil around red or white oaks has a correlation with the health condition of oak trees.  Besides the washing and Enrichment, the sample’s metadata is also very important for more insight into our research question.

Soil Washing and Enrichment for Sample (10.1.18):

Material:

•  15 ml Conical Tube
•  50 ml Conical Tube
• LB Broth
• Syringe Filter
• Vortex machine
• Centrifuge
• Micropipette
• Serological pipette

Procedure:

1. Set up an Aseptic zone
2. Placed sample Soil (10.1.18) to the 2 ml mark.
3. Poured LB Broth to the 12 ml mark.
4. Vortexed for 15 min to mobilize the soil.
5. Centrifuged at 10000 G for 10 min.
6. Filtered the supernatant with syringe filter to 10ml mark.
7. Added 0.5 ml Arthrobacter into the filtered supernatant and placed on shaker.

Soil Metadata for (10.1.18):

Soil Composition:

1. Added soil to the 4 ml mark on the falcon tube.
2. Added water to the 12 ml mark.
3. Added 3 drops of Soil Dispersion Liquid
4. Wait for 48 hrs to see the results

Soil pH:

1. Added a small amount of soil to the pH vial
2. Added water to 80% full
3. Shook the vial for 45 sec and waited for 2 min
4. put in a pH paper(should not exceed 1 inch)
5. Waited for 45 sec and compared the color with the pH scale to determine the pH of the soil
6. The pH level for Soil Sample (10.1.18) is  pH 6.0

Observations, Results & Data:

The plaque assay showed negative results and the control plate has four small distinct colonies, presumably contamination.

Interpretations & Conclusions:

There is still contamination on the control plate, however, the plaque assay still showed negative results, so the focus of the experiments will be moving on to the new sample(10.1.18).

Next Step:

In the next lab, I will be running plaque assay on sample (10.1.18)