Third Post!

While my time in the Young Lab has come to a close, I still have much to discuss. In the last few weeks I started on an actual experiment for the AzoBen UAA. I took agar plates, which are plates that bacteria grow on, and created an environment where bacteria can express the photoswitchable nature of GFP with my UAA. I took four plates, and to three of them I added 10 ml of agar, 10  μl of IPTG, 10  μl of Arabinose, 10  μl of chloramphenicol, 10 μl of Ampicillin, and 100 μl of AzoBen. On the fourth plate, I repeated the same procedure, except I did not add AzoBen. I then plated bacteria that could inherently grow GFP. The purpose of this setup, was to test whether I could detect changes in fluorescence when I irradiated the bacteria. Since three of the plates contained GFP with AzoBen, they theoretically should be able to change the fluorescence. I took one plate, irradiated half of it for 10 minutes, at 365nm. I then left the plate overnight in an incubator. When I came back and checked the fluorescence, I noticed that both sides were the same. This meant that either the experiment did not work, or that the plate did change, but changed back. Hoping that the result meant the latter, I took one plate and irradiated half of it for 10 minutes at 365nm, and checked it after an hour. To my joy, one side had a different fluorescence reading than the other. This meant that the experiment had work; but now I needed to check if the bacteria reverted back. After a few hours of hopeful checking, I saw that the irradiated bacteria were changing back to their original fluorescence! This was fantastic news, as I now have supporting evidence that my UAA is able to alter the function of GFP, but is also able to undo these changes.

[Read more…]

A Conclusion of Research, but not of Curiosity

As I write this final post, I cannot help but marvel at how quickly this summer seems to have passed by, and how much information I was able to gather within the past three months. I think mentioned in my an earlier blog post about the remarkable response rate to my survey, but even then I failed to anticipate the wealth of information I would receive from such a large sample size.

[Read more…]