I started off the day greeted by Dr. Bermek in front of the UNC Lineberger Cancer Center. She escorted me into the building and introduced me to others working in the lab. I got to work quickly and acclimated my Biotech skills with a DNA mini prep for three enzymes PUC19, PGLO, and P-Amalyse for the next years Biotech class. The protocol I used was striking similar to those I had seen in ADV Biotech. I suspended the DNA pellet in Buffer P1 using my well-practiced pipetting techniques and continued the protocol that called for a centrifuge. This piece of equipment I had used many times before, but it was interesting to learn something new about it. I learned that when setting the centrifuge to the correct speed I needed to use the g speed (e.x 17,500 x g) instead of the rpm (e.x 12,000 rpm) because each centrifuge has a different rpm but the g speed is the same. After completing the protocol Dr. Bermek and I used a Qubit Fluorimeter to check the concentration of the samples. The Qubit was a new piece of equipment I had never seen before, but it was simple, easy to use, and much smaller than the spec machine I had worked with in Biotech. Two of our samples, PGLO and PUC19 turned out well with a concentration around 50 ng/uL which was really awesome because Dr. Todd could keep them for next year’s class. Dr. Bermek and I could also use the PGLO for a future experiment. It was really nice that the first experiment I did at the Griffith Lab worked.
Next, I got to run some samples on a Agarose gel. I pipetted 6uL reactions on parafilm. Each reaction consisted of 1uL of the samples, 1uL of 6x dye, and 4uL of water. Once the 12 samples were pipetted onto the parafilm, Dr. Bermek and I loaded the samples into the gel box. I had run many gels in class before but I got to learn how to use and load a new system. Our gel also turned out successfully which was wonderful to hear, and the results we obtained are going to be helpful for future experiments. One of the coolest parts of my first day was getting to learn about the Electron Microscope or EM. The machine was very intimidating and marvelous at the same time. One of the people working in the lab was a major expert on the EM, and he was in the middle of fixing it when I arrived. To even be able to put samples in the EM there was a complicated process to prepare the samples that I got to help Dr. Bermek with. We used tiny grids (much smaller than a penny) that required special tweezer looking things to be able to handle them. We put solutions on parafilm and then dipped the grids into each one. Next, we placed the grids in one solution for 30 seconds and then quickly placed them in a different solution for 15 seconds one by one. To further prepare the grids, we used a complicated special machine that coated the grids in carbon and tungsten. I was easily impressed by the steaming liquid nitrogen used to cool down the machine. This process was intricate because the right amount of both carbon and tungsten had to be exact or we wouldn’t get proper results. I ended the day by looking at the images we obtained on the EM. I got to see single stranded, super coiled, and relaxed DNA up close which was magnificent. Overall, it was an action packed first day with lots of success. Working in a new lab was a little bit like the first day of Biotech class. There was a lot of new vocabulary, sciencey jargon, and math equations that were a bit challenging, but I got to learn so much on just my first day!
Working on my first experiment with Dr. Bermek.
The daunting Electron Microscope.