GFP sensors 2: Resistance is centri-futile

IMG_3469

Colorful tube of centrifuged GFP

IMG_3518

Exciting container of centrifuged dye

For these past two weeks, I have been growing bacteria to express fluorescent protein so I can harvest it. The protein expression process takes about one week, and I am making four different variants for my experiments. Fortunately, they can be expressed simultaneously. Unfortunately, some of the bacteria cultures have been refusing to grow for some reason. This has resulted in several weeks’ worth of many restarts of the whole process. Not knowing why the bacteria are failing to grow is especially frustrating because no one step can be corrected to fix the issue. This is particularly annoying because it is necessary to wait for a day to determine whether the bacteria grew or not. Despite these setbacks, it is important to have patience and try again even more carefully in hopes that something will work out.

This week, I finally have three of the four protein variants and can do experiments with them. Later this week, I will have results to compare and analyze. Honestly, one of my favorite parts of this project is using the software to make pretty graphs; however, it is necessary to have material to perform experiments on to get data to analyze. Additionally, it can get uncomfortable to do computer work in lab when the thermostat seems to be set to “finger-numbingly cold”.

For the next few weeks, I will work on expressing the fourth and final variant, which also happens to be the most promising in terms of results. I will also look into verifying that the dye and protein are both present and in the proper formation. I will also be trying to purify my product to get as little unreacted starting material as possible. Currently, large amounts of starting material are still present in the sample, interfering with my measurements. Hopefully, I can get these issues resolved and move forward with my project.