Additional note from Wednesday (8/30):

After further examination, it became very clear that there was Arthrobacter growth on our section of the top agar control plate. Unfortunately, this indicates that some amount of contamination must have occurred during the procedure. To avoid contamination in the future, taking care to keep instruments separate and avoiding any possibility of introducing contaminants to controls should rectify further problems with this.

Rationale: We obtained an additional sample of soil titled “Soil Sample B” in order to begin compiling data to answer our overarching research question.

Research Question: For Groups 3 and 4, we concluded that our research question to guide our soil collection would be old trees as compared with newer trees that needed to be transplanted or were recently planted. As a class, our idea is to come together after dividing into four groups to determine whether or not there are any trends or correlations based on the conclusions and data of each group that can guide us to a more definite question that the whole class can agree upon.

“Is there a significant difference in phage presence between old trees and new trees (that needed to be transplanted or were planted recently)?”

Procedure:

1. Class gathered to brainstorm and evaluate hypotheses.
2. Each table of two groups liked relatively different ideas, so each group went and began to collect soil to gather evidence from their own questions in order to develop correlations that could focus a class-wide study.
3. Groups 3 and 4 (making up one table group) concluded that the question we would begin to pursue was “does the amount of time a tree has been planted or if has been transplanted influence the phages in the surrounding soil?”
4. Therefore, group 3 found each found three separate wild oak trees and took 1/4 of a collection bag of soil from each tree along with one leaf. The bag was labelled “HMB Soil Sample B 9/5/18”.
5. The online metadata survey was taken to give additional context about the selected trees beyond the initial research question.
6. Soil in bag was brought back to the lab. 2mL of the sample from the bag was placed in a 15mL conical tube. The conical tube now containing soil, leaf, and excess soil were all placed in the bag and submitted to be placed in the refrigerator until Monday.

Metadata/Observations:

• Tree type: Live oak
• Tree Size: Approximately 15m tall, well greater than 200cm in girth.
• Canopy Size: Large, approximately 33m
• Health: Tree appeared to be very healthy. Branches all had leaves and the trunk showed very little evidence of decay.
• Soil: Moist, obtained from under grass next to roots.

***Pictures submitted via survey.

Conclusions/Next Steps: The dialogue surrounding the scientific question was constructive, and ideally, each group will be able to contribute correlation between their variables. The next lab procedure will involve washing the soil gathered today to obtain direct and enriched lysates that will be used to conduct a spot test and plaque assay. Depending on the results of the tests (spot and plaque assay), we will be able to begin to support or reject the research question surrounding transplant status and age of tree.