Luminescent Broth

In my two previous experiments, I was not able to successfully to isolate bioluminescent cells from a squid, so my Lab Supervisor Josh, has ordered Bioluminescent Bacteria for my next attempt at isolating bioluminescent cells. Meanwhile, I was asked to make media for culturing my bacteria.

It is important to provide the same environment and nutrition, for bacteria during culturing, which is found in its natural habitat. Therefore, the culture that I need to make must contain all the essential, nutritional components that these bacteria get in their natural habitat. A culture medium normally contains water, a carbon and nitrogen source, energy, trace elements and growth factors. In addition, the pH of the medium must be set accordingly. Liquid media are called “broth” so the first culture media that I need to make is “luminescent broth”.

 My media formulation:

Luminescent Broth
Peptone	10.0 g
NaCl	30.0 g
K2HPO2	2.0 g
MgSO4	0.25 g
Glycerol	2.0 g
Distilled H2O	1.0 L

Firstly, I weighed all of my substances using a scalar for accuracy. I placed the Erlenmeyer flask which contained 1 liter of distilled water and magnetic stir bar on the hot stir plates. Then, I turned on the plate to a medium temperature setting and started mixing at a medium-to-high speed while adding my substances one by one into the flask. It took approximately 10 minutes before all of the substances were dissolved.

Once all of substances were dissolved, I poured my solution into test tubes. I put caps on the tubes, in preparation for autoclaving, and placed a strip of autoclave tape on each of my test tube racks.

 

After that, I put my test tube racks into autoclave and I chose the media then hit start. This process took 46 minutes to complete.  After the autoclave cycle was finished, I took out the test tube racks from the autoclave chamber, and pushed all of test tube caps down.  I placed my test tubes containing the luminescent broth into the incubator. They will be stored to use later for culturing bioluminescent bacteria.