Rationale: perform spot test on direct and enriched isolations from the previous lab to check for presence of phages

Steps:

1. labeled plate with spots for enriched, direct, and negative control (phage buffer)
2. found enriched sample mass = 21.22g
3. centrifuged samples as a class
4. made LB agar for mine and Lilly’s plates, and a control plate

   1. Reagents	control plate	sample plates
      Arthro	NA	0.5 mL
      LB Broth	~2 mL	4 mL
      2x TA	2.5 mL	5 mL
      1M CaCl2	23 µL	45 µL

5. pipetted both solutions to mix
6. pipetted 5 mL control agar onto control plate
7. pipetted 5 mL of the arthro agar onto mine and Lily’s plates
8. Let plates sit for 10 minutes
9. Used a syringe and filter to filter enriched isolation into a new 15 mL conical tube
   1. filter had very little resistance – I tried two different filters and nothing changed
10. pipetted 5 µL of the enriched and direct isolations as well as the negative control onto the pre-marked places on the plates
11. let plates sit for 15 minutes
12. put plates into the incubator (NOT inverted)

Soil Metadata Note:

weight of dry sample and weigh boat = 6.31g

the soil level experiment that I had put into two different tubes had to be redone because it did not show anything. To improve the experiment I didn’t pour out the supernatant because that part ended up having a substantial amount of soil that shouldn’t have been separated from the rest of the sample.

Next Steps:

Check plates for sign of phages.

Perform plaque assay with enriched sample.

analyze new metadata results