Rationale

Begin soil metadata to determine percent water, pH, and type of soil collected. Soil washing and enrichment will be done in order to continue from testing through PCR.

Detailed Procedure: Soil Metadata

1. Weighed plate and added soil. Recorded both plate and plate +soil
2. Filled soil up to 10 mL mark in a 50 mL vial and filled rest with D.I water up to 30 mL mark.
3. Added 3 drops of soil dispenser solution and shook for 30 secs
4. Left to rest on rack for 48 hours.
5. Pinched soil into pH vial and filled rest with D.I water and shook for 10 secs
6. Left to rest for 2 mins
7. Pinched off 1 inch of pH paper and dipped into pH vial for 45 secs.
8. Recorded pH of soil

Detailed Procedure: Enrichment

1. Filled soil in 15 mL vial up to 2 mL mark
2. Filled LB Broth into vial from Lab Day 14 up to 12 mL mark in aseptic zone
3. Shook using vortex for 10 mins.
4. Weighed vials and centrifuged for 5 mins
5. Took dropper and poured supernatant into 3 separate wells around halfway.
6. Heated up supernatant at 55 degrees for 5 mins and 60 degrees for 5 mins with shaker-thermostat
7. Took 100 microliters of Arthro into 2 of the 3 wells.
8. Incubated it and shook in the machine.

Results metadata

• pH= 6.5

Conclusion/Next Steps

Group members decided to heat up the supernatant to kill of any bacteria and let it cool down before adding the Arthro. This was a experimental way of making enriched lysate without manually filtering the supernatant. In the next lab day, group members will run PCR.