Lab Proceedings

Day 1:
We mixed and poured the Agar base into petri dishes and left them to set overnight.

Day 2:
We rehydrated the E. Coli in two batches; one with the bioluminescent protein and one without. We then poured bacteria onto plates with 3 different mediums, standard LB-Agar, LB-Agar with ampicillin, and LB-Agar with ampicillin and arabinose. This left us with 4 different plates (top left image)

Day 3:
Came in and checked on progress, very little growth apparent, left bacteria in incubator (top right image)

Day 4:
Results: (bottom images)

Agar and Bacteria Type	Expected Results	Our Results
+pGLO LB/amp	~75 white colonies	No growth
+pGLO LB/amp/ara	~75 fluorescent colonies	3 fluorescent colonies
-pGLO LB/amp	No growth	No growth
-pGLO LB	Even lawn of off-white bacteria	Even lawn of off-white bacteria

At this point, we were unhappy with our results. We did not achieve the growth we expected, and decided to conduct our own follow up experiment using the fluorescent colonies we had. To allow the fluorescent bacteria to proliferate, we decided to incubate the sample for a longer period for more dramatic results.

Day 5:
The fluorescent colonies were much larger than before, but there were additional, non-fluorescent colonies that somehow contaminated the sample. We decided to seed a new plate with bacteria from one of the fluorescent colonies in the shape of SJP in the LB/amp/ara. We left it in the incubator. We also applied the fluorescent bacteria to all three types of Agar dishes, to test survivability and to see if they retained the fluorescent trait.

Day 6:
The lettering was not perfect, and looked more like SJD than SJP, but the letters were fluorescent and were easily recognized. The two samples on the LB and LB/amp plates survived, but did not retain their fluorescence. The sample on the LB/amp/ara plate grew to an even lawn of fluorescent bacteria. The colonies on the original plate grew slightly larger, as did the contaminant colonies. (see picture below)​

Conclusion:
While the bacteria did not grow as much as expected, or at all in one case, and there was an issue of contamination, the lab went well and we were not only able to successfully produce the fluorescent bacteria, but also to design with it and run another experiment of our own.

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