Rationale

Today we will collect soil, wash it, and collect metadata.

Procedure

• Established an aspectic zone.
• With new soil collected, 2 mL of soil sample #4 and 8 mL of LB Broth was added to a tube. The tube was then vortexed for 10 minutes and massed.
• After 10 minutes, the vortexed soil was centrifuged at 5000g for 10 minutes. The supernatant produced was filtered with a syringe filter into another vial and 0.5 mL of Arthrobacter was added. The vial was incubated for 48 hours.
• To determine the percent water of the soil, a weigh boat was massed and then massed again after ~3g of soil was added. It was set under the fume hood for 48 hours to dry.
• To determine the percent sand/silt/and clay of the soil, 10 mL of soil was placed into a Falcon tube and was filled to the 30 mL mark with D.I. water. 3 drops of soil dispersion liquid were added and the tube was vigorously shaken for 45 seconds. It was then set under the fume hood to sit for 48 hours.
• To measure the pH of the soil, 1 mL of soil was placed into a pH vial. It was then filled to the top with D.I. water and was shaken for 30 seconds. The vial was set aside for 10 minutes. After 10 minutes, pH paper was placed in and the pH was measured.

Observations

Mass of the weigh boat: 2.328 g

Mass of the weigh boat and soil: 5.985 g

pH of the soil: 6

After the tube was centrifuged, the supernatant produced was clear and slightly yellow in tint. 9 mL of enriched lysate was able to be produced.

Conclusion/Next Steps

Using the enriched lysate from the soil sample, a plaque assay and a spot test will be conducted to test for phage presence. The results of metadata collection will be observed as well.