Transformation Tuesday

Although the phrase, “Transformation Tuesday” is typically reserved for hashtags on Instagram and Twitter (and Neville Longbottom), it also applies to my experiences of the past week in lab.

In fact, one week ago, on Tuesday, I was completing a bacterial transformation. That was also the day that my lab experience “transformed” into my own, unique project! While my work is only a small, small part of the lab’s overall project, I am thrilled to be contributing to exciting and innovative scientific discoveries!

My project in a nutshell: Right now, the lab is seeking to analyze the interactions of every human protein with histones. We are using protein arrays (and lots of other REALLY cool lab protocols) to identify particular protein interactions. As such, I have been tasked with purifying and (eventually) identifying the interactions among two groups of proteins: orca and orc1.

In the past 7 days, I have gained a sense of self-confidence that I never expected from this experience. I guess you could say that the training wheels are off. Going in, I thought that I would feel lost and confused all summer. Who knew there was so much math in science?!

Thank goodness for my colleagues and mentors in the lab! They have made the transition from the classroom to the lab an enjoyable and exciting experience.

So, here’s a run-down of the past seven days:

1. Bacterial Transformation (1 day): I performed a bacterial transformation with the goal of modifying E. coli that would incorporate my target proteins (orca and orc1) into their genome. Below is a picture of the orc1 protein. I am specifically working with orc1 BAH 185 and BAH 275.

2. Induction Tests (2 days): I have been doing a series of tests to see if a different strain of E. coli would produce my target proteins for me. I’m only about halfway done with these, but so far, the bacteria are producing at least one of my proteins–yippee! This process consists of a lot of sampling, then waiting, then sampling and waiting some more. Then, once you have bacterial samples from 1) pre-induced, 2) after 1 hour of induction, 3) after 3 hours, and 4) overnight, you can compare the samples to see if the protein was produced. The goal is to see a relative increase in protein production over time. The way that this test is visualized is by running each sample on a polyacrylamide gel and comparing the “pre-induced” to the “overnight” sample.

3. Coomassie staining (1 day): Funny word, right? I’m fairly certain that this is a procedure that I’ve done before with my AP biology students. However, when we order the kit with the stain, the company calls it CarolinaBlu stain–alas… Essentially, it is a blue dye that attaches itself to any proteins that passed through the gel (see step 2). After the gel has been dyed, it is destained using white vinegar (acetic acid), so that only the bands remain. Here’s a picture of what mine looked like!

Right now I’m repeating steps 2 and 3 with a different set of proteins. I am hoping that by the end of the week, I can scale up the procedure and get the bacteria to make my proteins in large batches!

Again, this week has been a true transformation: learning new techniques, working independently, and finally feeling confident that a) I’m contributing to the lab and b) that I know what I’m doing!