Rationale:

For this lab, a plaque assay will be created, the % soil composition will be determined, the enriched lysate will be filtered, and % water experiment will be set up.

Procedure:

First, the lab table was wiped down with CiDecon and Ethanol, after which an aseptic zone was set up to prevent contamination. The soil composition was determined by removing the water from the top. After, the procedure to determine % water was set up by weighing a weigh boat along with 3-5 g of soil. 1.2 mL of enriched lysate was added to a tube then put into the centrifuge for 10 minutes to be filtered. After 10 minutes, the lysate was filtered using a syringe and filter. Then, 0.5 mL of Arthrobacter and 1 µL of lysate were mixed together and left alone for 10 minutes. Next, LB broth (2 mL) and CaCl2 (22.5 µL) were combined. After 10 minutes, the 2x Top Agar (2.5 mL) was added to the top agar solution along with the lysate and Arthrobacter. The solution was then poured onto the plate and given enough time to solidify. After solidification, the plates were placed into the incubator and left there for 48 hours.

Results and Analysis:

% Soil Composition 

Out of 4 mL

% Soil (.75 mL)= (.75/4) x 100= 18.75%

% Sand (2.75)= (2.75/4) x 100= 68.75%

% Clay (.5)= (.5/4) x 100= 12.5%

Rather than having a smooth layer, the control plate had a bumpy texture.

Conclusions:

First, the tables were cleaned with CiDecon and ethanol along with an aseptic zone. The soil composition was determined then a % water procedure was set up. 1.2 mL of lysate was centrifuged for 10 minutes then taken out and filtered using a syringe and filter. Then, the lysate and Arthrobacter were mixed together to infect. The LB broth and CaCl2 were added together into a vial of which would become the top agar solution. After 1o minutes, the 2X top agar was added along with the lysate and Arthrobacter onto the plate. It was left alone to solidify then put into the incubator.

Future Plans:

The plaque assays will be checked for the presence for plaques. If there are no plaques with contamination, another plaque assay will be created. If there are no plaques and no contamination, then a new soil sample will be collected, If there are plaques with no contamination, a plaque will be picked and will be diluted using phage buffer. After diluting it, a plaque assay will be created to begin the process to get a high concentration of plaque.