Week 10 already? What a great summer!

This week I tried to get some more answers from the mutants that I didn’t send to be sequenced last week. To prepare for Tuesday’s transformations, on Monday I made media and inoculated all of my remaining mutants and two controls for overnight cultures. Since Monday was just waiting for bacteria to grow, I had time to work on my final paper and presentation. My mentor offered to listen to my presentation and gave me some feedback. This was really helpful and also very kind since she has to hear it a few times anyways!

Tuesday morning I made all of the remaining mutants competent the “quick and dirty” way, but there was nothing quick about it. I did two transformations for each of the 20 mutants… which is 40 transformations! Once I had made my mutants competent, I transformed them with two different plasmids: one known to lack flhF and the other lacking fleN. These transformations were a way to test if the initial transposon insertions in these mutants were in one of these genes, instead of sending them to be sequenced.

I gave my final presentation at the Tebo Lab meeting. Preparing a presentation was a nice way to conclude my work here this summer because it makes me think back and reflect on the past ten weeks. Of course it’s also a little nerve-racking giving a presentation to a room full of people with Ph.D.’s in the topic… It went just fine and I got a few questions, so my audience must have been engaged and interested in my project.

On Wednesday I made a lot of media… surprise surprise! My mentor said it would probably be the last batch of plates I’d pour this summer…which is crazy to think. I remember pouring plates for this first time this summer and being so nervous I would spill that my hands were shaking, and now it’s just so routine.

Thank goodness all of my transformations worked. All of the LBgentamycin transformation plates had plenty of growth on them, which means that many of the colonies took up the plasmid as I wanted. Since the next step in this process is to screen for oxidation, I had to streak out a colony from each transformation plate onto a lept plate so oxidation could begin.

By Thursday, the mutants I plated on Lept had not begun oxidizing, so I had to be patient. My mentor read over my final paper and gave me a lot of really helpful revisions and suggestions. The biggest struggle with this paper is deciding how much detail to include, because I’ve done so much this summer and need to sum it all up in a few pages!

Wow. I can’t believe ten weeks have already gone by. This has been such an awesome experience and I couldn’t thank my mentor enough for teaching me so much this summer. As a little gesture to show my appreciation, on Friday I poured four batches of plates for her that she’s going to need for next week.

Only a few mutants were oxidizing by Friday, but they weren’t oxidizing with the plasmids that I had expected…which is really cool! This might suggest that the transposon insertion was in a gene unrelated to flagella synthesis! My mentor wants to map the transposon insertions for these mutants that became of interest.

My mentor and a few other lab mates treated me and the other interns to lunch on Friday. It was so nice of them to do that, and really made me feel like I’ve become part of the Tebo lab. I’m going to miss seeing this group of people every day!