Results:

From the plaque assay ran on Friday (10/12), 4 plaques were found as shown below.

Rationale:

Since plaques did appear, soil metadata will be collected for future correlations. Also, a plaque will be picked and serial dilutions will be made to be ready to plate to start the quest to find a high titer.

Procedure:

1. Once an aseptic zone was established, about 3 grams of soil E was poured out into a weigh boat labeled “KEA 10/15 soil E” and left under the flume hood to dry out.
2. A falcon tube labeled “KEA 10/15 soil E” was filled with 10 mL of soil E, 20 mL of de-ionized (DI) water, and three drops of texture dispersion liquid.
3. The “KEA 10/15 soil E” falcon tube was shaken for 30 seconds and then left under the flume hood for the next 48 hours.
4. A small amount of soil E was placed in a pH vial and the rest was filled with DI water.
5. The pH vial was shaken for 10 seconds and then settled for 2 minutes.
6. Used pH paper to determine the pH of the soil.
7. 100 µL of phage buffer was placed into a microcentrifuge tube labeled “KEA 10/15 10⁰.”
8. Used a micropipette tip to touch the plaque and then swirled the tip in the “KEA 10/15 10⁰” microcentrifuge tube.
9. The “KEA 10/15 10⁰” microcentrifuge tube was vortexed.
10. 10 µL of “KEA 10/15 10⁰” was added with 90 µL of phage buffer to create 10^-1
11. 10 µL of “KEA 10/15 10^-1” was added with 90 µL of phage buffer to create 10^-2

Observations:

• The 4 plaques found on the “KEA 10/12/18 PA Soil E” plate were small and distinct.
• The weigh boat weighed 2.39 grams. The weigh boat with wet soil weighed 6.15 grams.
• The pH of the soil was determined to be 6.0 as shown in the picture below.

• Dilutions were created all the way out to 10^-7.

Next Steps:

On Wednesday, the dilutions will be plated.