TRAIN Keeps Rolling

 Introduction: TRAIN Keeps Rolling

    Week 2 of research was getting to the nitty-gritty. Since introductions were out of the way already, the team and I got right to it. The question arose from our mentor, Jonathan, "how does bacteria spread"? As my mask covered the blank expression across my face 😐, I knew I needed to say something. As the table sat quiet, a spark of genius came across my mind, "something touching it then touching something else" I blurted. While in my mind nobody could tell me I am wrong, I was only partially right. Jonathan then explains to the team and me, that there are four ways bacteria can spread; conjugation, transformation, transduction, and vesiduction. For our research, we will be focusing on transformation, which is when genetic material is directly taken up and incorporated by a cell through the cell membrane. 

Magnetic Pull: Gel Electrophoresis

    After Jonathan went into great detail about the different methods of how bacteria spreads. It was now time to start experimenting. The term gel electrophoresis is then brought to light as Jonathan brought out a contraption one could only assume is used for shocking bugs. This plastic contraption has a red, positive terminal on one side, and a black, negative terminal on the opposite end. There was a grayish, wet-looking gel that was residing in the middle of the contraption, almost like a waterbed for the bugs I was assuming we were shocking. Jonathan then explains what this contraption is, and what it does. You place samples inside of wells on the gel, and the magnetic pull from a positive terminal will then separate the samples. Sounds simple enough. There are steps you have to take before shocking the gel. First, we need to add TAE as a buffer, and then verify that the gel is covered completely by the TAE buffer. Second, this should probably be the first step, but make sure the gel is oriented the correct way. We want the long end facing towards the positive terminal, otherwise, the samples will be pulled right out of the gel if the short end is facing the positive terminal. Third, we GENTLY add our sample to the wells. There were five wells inside the gel we used, 2 ladders, and 3 samples. Seeing as the gel was oriented the correct way, and the wells had the samples, Jonathan yelled "BRING OUT THE TRANSFORMER"! Almost like something you used to type on in the early '90s, the transformer was at the table. The next steps are simple. make sure red is too red, and black is too black, just like when you jump start a car. Run constant amperage at 75 mA, hit the run button, and verify it is working from the air bubbles coming from the terminals. 

My Time to Shine: LB Plates

    As I mentioned in my last blog, we will be using two LB plates, one without ampicillin, and one with ampicillin. We would also be using TGY plates, one with kanamycin, and one without kanamycin. Visually we could tell there was growth on three out of the four plates. There was growth on the LB plate with and without ampicillin, and growth on the TGY plate without kanamycin. As a team, we discussed how we could verify the data at hand, simply put, we use more samples to verify the data. This was a test batch, mainly for the team to practice our infamous four-quadrant method. Which we all could still use practice. The top left plate is the TGY without kanamycin, the top right is the TGY with kanamycin. As we can see, there is little growth in the top left, but no growth in the top right. The bottom left (which is expertly done by myself 😉) is the LB without ampicillin, and the dish on the bottom right is LB with ampicillin. As we can see, both dishes have bacteria growth. 

 

Summary

    This was a great first full week of research! Since it was not my first day anymore I was not completely lost like I was the previous week. Our team worked great together, and we (probably just me) learned a lot. I can say my favorite part is the practical, hands-on stuff. I like getting in there and learning through trial and error, which might be counterproductive in a lab setting. I learned a lot, and just like the week prior, I am excited to keep learning more.