Bioprinter update 5

After obtaining a high titer lysate for one of my M.smegmatis phages, I have spent a large portion of my time in lab trying to prepare electrocompetent cells. Once I have my cells ready for electroporation, the next step will be to insert my shuttle vector into them. Electrocompetent cells are also essential if I ever want to begin engineering phage, a potential direction that I could pursue over the school year.

I was able to utilize my bioprinter for several spot tests with my smegmatis phage. After calculating my titer, the concentration or phage forming units per milliliter, I was able to calculate how many theoretical plaques I would have per pipetted volume. I created a series of serial dilutions from my lysate and plated up to a 10^-7 dilution.

In order to examine phage infection dynamics with biofilms, I am in the middle of a series of experiments in which I have infected biofilms with phage at different time points. At the end of a four day period the biofilm will be stained with crystal violet for visualization.

 

Spot Test