Rationale

Perform plaque assay from Soil D without any trace of contamination.  Two members from group 6 did spot test while one did plaque assay. Each test had an individual control plate.

Detailed Procedure

1. Syringe filtered enriched lysate.
2. Took 10 microliters of filtered enriched lysate and added to 0.5 mL Arthro. Sat for 15 mins.
3. During 15 mins, took 4.0 mL LB Broth, 45 microliters of 1M CaCl2, and 5.0 mL of 2X TA into separate 50 mL vial. Mixed through pipetting.
4. Used sterile pipette and took half of solution from step 3 into another separate 15 mL vial. Labeled as control.
5. After 15 mins were done, took solution from step 2 and mixed through pipetting into previous 50 mL vial.
6. Took both vials into water bath and waited for plate to be ready.
7. Labeled plates as “Plaque Assay” and Plaque Assay Control.”
8. Took control vial and poured into control plate and same with other plate+vial.
9. Waited 15 mins to solidify.

Conclusion/observations

In the beginning, the unfiltered enriched lysate had arthro all over the tube which was because the vial was not taken out during the past 4 days. Due to more experience, prevention of contamination technique were better (accidental spills, not being in aseptic zone.) Control plate had no bubbles and did not have problems sliding. Experiment plate had two bubbles and continued to slide even after 15 mins were done. Both plates could not be inverted in the incubator. In the next lab day, future steps would be to check for any signs of plaque. Since within the group, both spot test and plaque assay was performed, so if no plagues were found, then a new soil will be tested next. if there is time this week.