8/29/18

Spot Test Soil A Results and Plaque Assay Soil A

Rational:

To confirm the results of the spot test on soil A with a plaque assay of soil A. This will give results if the sample was contaminted and will confirm results found in the spot test.

Procedure:

• Cleaned the lab desk with CiDecon and 70% ethanol
• Labeled the plate plaque assay soil A with enriched
• Labeled conical vial team 2 TA
• Added 22.5 ML CaCl2 using a micropipette
• LB broth splashed out preventing us from knowing how much was added- conical vial was thrown out
• Labeled new vial team 2 TA
• Added 22.5 ML CaCl2 using micropipette
• Added 2.0 mL LB broth using a serological pipette
• In order to have 5 mL for each team member from the vial and 1 mL for the control plate we added 67.5 ML more CaCl2 with a micropipette and 6 mL LB broth with 10 mL pipette
• Labeled tube with arthrobacter (0.5 mL arthrobacter) arthrobacter with enriched soil A
• Added 10 ML lysate to the tube with a micropipette
• Set a timer for 15 min
• Added 10 mL of TA
• Added 5 mLmof TA mixture to the tube with the lysate (some spilled) (KEA and my tubes were mixed up so the plates were re-labeled)
• Added 1 mL of TA mixture without lysate to the control plate
• Put plates in the incubator at 26 C at 4:00 8/29
• Cleaned lab desk with CiDecon and ethanol

Results:

*results from spot test on 8/27                                                                                                                                   Contamination was found on the control plate as well as my plate so plate                           

fig.4 – shows where contamination and bubbles                  fig.7 – shows the plate after incubating for 46 hrs.               are found on the plate

fig.5 – shows where contamination was found                     fig.6 -show the dots of contamination on the control plate  on the control plate

Analysis and Interpretation:

The results from the spot test indicated that there was no bacteriophage present in the soil A, however the contamination observed prevents this from being confirmed in the spot test.

Future Plans:

I plan to check the plaque assay for plaques during the next lab. If no plaques are present then a new soil sample will be collected. The new soil samplewill then have LB broth added and filtered. Then a new spot test and plaque test will be done (not all in one lab). If a plaque is observed the plate will be saved and a new test to confirm the presence will be done. If there is contamination the test will be done over trying to prevent contamination again.