Phage Lab Report

Collecting an Environmental Sample: A soil sample was obtained from the greenhouse in Abelson Hall on the seventh floor at Washington State University. The coordinates of the site are 46.729584, -117.165361. The sample was wet, and the temperature was 24 degrees Celsius.
Isolating a novel phage from the environment: The purpose of this experiment was to extract phage from the sample to infect the host bacteria. This protocol shows the variety of phages in the sample. 3-5ml of the sample was poured into the bioreactor enrichment medium. Next, .5ml of M. foliorum was poured into the bioreactor, closed, labeled, and put in a shaker for 48 hours.
Enriched Isolations: Enriched isolation was used to amplify phages present in the sample. This protocol allows bacteria to grow, replicate, infect, and form a large mass of phage. 1ml of the enrichment and soil mixture was transferred to a microfuge

This procedure requires plaque isolation, which enables characterization of a single phage population. A single plaque was chosen from the plate by circling the plaque with a sharpie and touching it with a sterile wooden applicator. The plaque was aseptically streaked onto a new agar plate. The wooden applicator was disposed, and a new applicator was obtained to start a new streak, starting from the end of the previous streak and repeated for a total of 3 streaks on the plate. The end of the third streak was marked with an “X” using a sharpie. The plate was then properly labeled. To prepare the agar plates, 20ul of CaCl2 was added to a tube of top agar. Next, a tube of top agar was poured into a tube of M. foliorum and mixed gently. The mixture in the last step was then poured onto the L-agar plate where the “X” mark was made. After sitting for 10 minutes, the plate was incubated in the 30-degree Celsius incubator for 48 hours. The experiment will continue next lab by performing pure phage