Title: New Soil Enrichment + Metadata

Date: September 10, 2018

Rationale: The new soil collected in the previous week is to be washed and enriched for further testing to make progress on our research question

Procedure: Aseptic zone created by the following procedure:

• Counter washed and wiped with CiDecon
• Counter sprayed with 70% EtOH and allowed to evaporate completely (to dehydrate and kill any bacteria on the counter and avoid contamination)
• Ethanol lamp lit to create rising heat and a current that protests samples from falling contamination.

1. ~2mL of soil was added to a 15mL vial
2. ~10mL of LB Broth was added to bring the vial up to the 12mL mark
3. Vial was shaken for 10 minutes

Meanwhile, soil metadata was taken

% Water: An empty weigh boat was massed at 2.39 grams, soil was added to bring the mass of the weigh boat to 7.76 grams. The weigh boat was left to evaporate for 48 hours. The mass of the dry dirt will then be used to calculate the % water of the soil.

Sand, Silt, Clay: ~4mL of dirt added to Falcon tube, DI water was added to cover the dirt, and soil dispersion liquid was added to separate the soil. The entire mixture was shaken for 30 seconds and left to disperse over a 48-hour period.

pH: A small amount of dirt was added to a pH testing vial and was filled the rest of the way with DI water. The mixture was shaken for 10 seconds and the pH was recorded on a pH testing strip. The recorded pH was 6.0

A 22 micrometer Top filter was then used to generate ~7.5 mL of lysate. 0.5mL of Arthrobacter was added to enrich the lysate and incubated at 28 degrees Celsius for 48 hours.