October
19
10/15/18- Plaque Assay Sample C
10/15/18
Objective:
- To make a plaque assay to test for the presence of phages in soil sample C
Pre- Lab Observations and Data:
The metadata for soil sample was collected from the results of the procedures performed on 10/10/18
% water = mass of water/mass of soil and water x 100= 1.038/5.095 x 100= 20.37%
pH of Soil= 6
%sand= 9.1/10 x 100= 91%
%silt= 0.4/10 x 100=4%
%clay=0.5/10 x 100=5%
Procedure:
- The aseptic zone was set up.
- Some of the enriched lysate prepared on 10/10/18 was filtered into a microcentrifuge tube.
- 0.5 ml arthrobacter was retrieved and enriched by filtered lysate.
- The vial was then allowed to rest on the test tube rack for 15 minutes
- While the vial was resting, one Top Agar mixture was made for the group.
- 8 ml of LB broth was transferred to a conical vial.
- 90 microliters of the CaCl2 was transferred to the conical vial with the LB broth.
- The vial was then set on the rack.
- 10 ml of the 2X TA was added to the LB broth and Cacl2 after the sample was allowed to enrich the lysate for 15 minutes.
- 4.5 ml of the top agar mixture was transferred to the test tube with the enriched lysate.
- The contents of the test tube were then poured onto the agar plate.
- Part of the top agar mixture was poured into the top agar control plate for the group.
- To let the top agar solidify, the plates were allowed to rest for 20 minutes.
- The plates were placed upside down in the incubator, where they will remain for 48 hours.
Analysis:
analysis of the soil metadata leads to the conclusion that the soil that was collected loamy sandy