Rationale: Wash new soil samples. The lab was out of filters so we designed a protocol to heat and kill the bacteria.

Process:

1. ~3 mL of soil and ~9 mL LB broth were added to tube and vortexed and shaken for 10 minutes
2. the mixture was centrifuged for 5 minutes at 3000 G

Heating and Killing Bacteria Method

1. A hot plate was heated to 55° C
2. Using a transfer pipette, supernatant from the centrifuged lysate was added to the wells of a plate (3 wells per lysate)
3. The sample plate was heated for 5 minutes
4. The hot plate was turned up to 60° C
5. The sample plate was left to heat for about 15 minutes
6. The sample plate was taken out to cool and then 100 µL of arthro was added to 2 wells for every 3 belonging to 1 lysate
   1. The sample plate MAY not have cooled down enough
7. The hot plate was turned down to 27° C and set to shaking at 150 RPM
   1. The sample plate may also have been returned to the hot plate before the hot plate had been cooled sufficiently
8. The sample plate was to left to incubate until the next class

Soil Metadata

• mass of weigh boat = 2.315 g
• mass of weigh boat and wet soil = 14.225 g
• soil sample was left under hood until next class

Next Steps

plate the enriched isolation to test if the heating and killing bacteria method was successful. In addition, wash soil samples again with filters.