October 19

OCTOBER 12TH, 15TH, AND 17TH- Labs

OCTOBER 12TH, 2018
- RESULTS: (RESULTS FROM 10/10 WERE VIEWED UNTIL 10/12)
  - The results showed in Figure 17 show a small amount of contamination on the arthro control, and TA control and the plaque assay were clear of any contamination
  - The plaque assay was negative for phage presence
- CONCLUSION:
  - The contamination seen on the arthro control is most likely from contamination from a airborne bacteria that ended up on the plate. Therefore we know the arthro was not contaminated, and it can be concluded that this soil sample does not contain phage.
- OBJECTIVE:
  - Enrich new soil sample
- PROCEDURE:
  - The sample was retrieved from the incubator when it was noticed that the sample was pink due to contamination that can be seen in Figure 18
  - Then the sample was placed into the centrifuge for 10 minutes at 3,000G
  - The bacteria then gathered at the bottom of the tube as seen in Figure 19
- RESULTS:
  - Due to contamination the sample could not be used
- CONCLUSION:
  - The sample was discarded due to the contamination having affecting the potential phage population, if there was phage present.
- NEXT STEPS:
  - Wash and enrich the sample soil sample
OCTOBER 15TH, 2018
- OBJECTIVE:
  - To wash the soil sample and have no contamination
- PROCEDURE:
  - Soil was filled to the 2mL mark of a test tube
  - Then 10mL of LB broth was added to the tube
  - It was shaken fro 15 minutes, before then being placed into the centrifuge for 10 minutes where it was then spun at 3,000G
  - One the tube being centrifuged was done, a syringe and .22 𝝁m sister was then used to filter out the supernatant
    - - Then .5mL of Arthrobactor was added to the filtered supernatant
      - The tube was then left in the incubator
- RESULTS:
  - No results to report
- CONCLUSION:
  - No results to discuss
- NEXT STEPS:
  - Conduct PCR
OCTOBER 17TH, 2018
- OBJECTIVE
  - conduct PCR, and understand the fundaments behind it
- PROCEDURE:
  - Lysate was spun in the centrifuge for 5 minutes at 3,000G
  - 2mL of the spun lysate was put into a micro centrifuge tube and was boiled
  - After the sample was boiled
  - Four tubes each containing 12.5𝝁L Taq + dNTPS were then labeled “1” “2” and “3” and “c”
  - The tubes labeled 1,2, and 3 all had 1𝝁L of DNA added from each soil sample (samples 5L and 5S)
  - The tube labeled c (serving as the control) then had 1𝝁L of dd water added to it
  - 4 𝝁L of the corresponding primer was added to tubes 1,2, and 3 (ex. primer P1 was added to tubes 1)
  - 4 𝝁L of P2 was added to the control tube
  - Then 6.5 𝝁L of dd water was added to each tube
  - The sample was then taken to be run through the PCR machine by T.A
- RESULTS:
  - Waiting on results
- CONCLUSION
  - Waiting on results