Title: Soil Sample 3 Enrichment & Metadata

Date: 17 October 2018

Rationale: A 3rd soil sample was collected from a planted tree in front of Earle Hall, the soil will be washed and enriched as well as the metadata of the soil taken. The resulting lysate will be incubated and tested in the following week.

Procedure: Aseptic zone created by washing lab bench with CiDecon and Ethanol, and by lighting an ethanol lamp

• 2 mL of collected dirt added to 15 mL conical vial
• LB Broth added to ~12 mL mark
• Shaken/vortexed for 10 minutes
• Centrifuged for 5 min @ 3000 G
• Filtered through 22 micron Top Filter
• 0.5 mL of Arthrobacter added to ~7.5 mL of filtered lysate
• Placed into shake incubator

The metadata of the soil was also taken with the following procedure:

• % Water:  a small amount of soil was placed into a weigh boat and massed at 11.26 g, the soil will be left to evaporate and will be weighed after evaporation, the % water composition will then be calculated
• % Sand, Silt, Clay: ~4 mL of dirt added to falcon tube, DI water added to 12 mL mark on tube. 3 drops of Soil Dispersion Liquid was added, the mixture was shaken for 30 seconds, and the mixture was placed under the fume hood to disperse.
• pH: A small amount of dirt was added to a small pH vial with DI water and shaken for 10 seconds. A pH strip was then inserted into the mixture and the pH was then recorded.

Conclusions: With the new soil sample, another attempt at finding phage can be run. A spot test and a plaque assay will both be done to test phage presence, and picked if positive results are observed. The pH of the soil was recorded at 6.5 and the rest of the metadata will be recorded in the next entry.