10/11/18

Objective:

• To extract lysate from soil sample C.
• To collect soil metadata on soil sample C

Pre-Lab Observations:

The control plate was not contaminated. there were no plaques on the plaque assay form 10/09/18. It seems that there were no phages in soil sample B and therefore a new sample will be analyzed.

Procedure:

1.  An Aseptic zone was set up.

Washing and Enrichment:

1. Using a serological pipette,  10ml of LB broth to a 15 ml conical vial containing 2 ml of soil sample C.
2. The vial was then shaken and vortexed for 15 minutes.
3. after  15 minutes, the vial was weighed and another vial of water with a similar mass ( within 0.01) was prepared and they were both centrifuged for 10 minutes.
4. using a sterile syringe and a syringe tip filter (22 microns), 10 ml of filtered lysate to a conical vial.
5.  0.5 ml of arthrobacter was added to the vial and labeled as enriched sample
6. the tube was placed in the shaking incubator for 48 hours
7.  rest of the lysate was filtered into another conical vial and labeled as direct isolation.
8. store direct isolate in the fridge

% Water

1. weighing boat was weighed on the scale ( g)
2. some soil was added onto the boat and weighed
3. the difference of the final weight ( boat and soil ) and initial weight (boat) was taken to find the weight of the soil.
4. the soil was placed in the fume hood for 48 hours, after which it will be weighed again.

Sand, Silt, Clay

1. 10 ml of soil sample was transferred to a falcon tube.
2. 20 ml of DI water was added to the tube along with soil dispersion liquid
3. the vial was shaken for 30 seconds and allowed to rest for 48 hours.

pH

1. a small amount of soil was added to the pH vial
2.  water was added to the vial until the level reached the top of the vial
3. the vial was shaken for 10 seconds and then allowed to rest for 120 seconds
4.  after 120 seconds, the pH paper was immersed in the vial for 45 seconds
5. the pH was determine using the color scale on the chart in the pH paper dispenser.

Analysis and Conclusion

due to the lack of legitimate plaques in the past few plaques, it is probable that phages were not present in soil sample B and it was time to move on. soil sample C was collected from a tree in close proximity from soil sample B, the possibility of correlation of presence of phages to the proximity of soil where phages are absent is logically nonexistent, this sample may lead to reveal a possibility if phages are absent, leading to another possible question to answer in conjecture to this research currently being performed.