Doug Reinisch, Week 1

My first week at Drexel was both challenging and relaxing at different points.  Travelling to an unfamiliar part in Philadelphia, one of the biggest cities in America is not an easy feat.  It took me a couple of rides on the PATCO and wandering around 15th street to finally figure out how to have a smooth commute.

My first day, Monday June 19, was my longest and most tedious, being jet-lagged and recovering from a slight caffeine addiction thanks to the China trip didn’t help either.  Within ten minutes of my being in the lab, I was already watching my first rat surgery which was conducted by a graduate student (Nick).  The goal of the surgery was to isolate the hepatocyte cells from the rest of the liver.  Afterwards I began a series of introductions to different laboratory members and filled out a series of paperwork.  My PI, Dr. Bouchard then gave me and another high school student Kennadi, who started work on the same day, a personal tour of his lab as well as the rest of the floor we work on.  After being introduced to the cafeteria and having lunch with a different graduate student, I was finally introduced to Joe, the graduate student who I will be working with the remainder of the summer.  He gave Kennadi and I some brief background information on HBV and HBx (most of which I already knew from EXP and AP Bio) before our lab meeting that afternoon.  At the lab meeting, Joe gave the presentation about his former, current and future research while Dr. Stills and Dr. Bouchard gave him a series of questions to ensure what he is researching shows promise.  After all this, I still had an hour and 15 minute commute home as the thunderstorms rolled in.

My home for the next 6 weeks!

Luckily the following day was not so long nor hectic, allowing me to recover and prepare for the rest of the week. On Tuesday both Joe and Dr. Bouchard introduced me to the specifics of Joe’s project, that I will be assisting him with.  Joe’s mutagenesis project has a goal of finding where on the genome of 154 amino acids causes HBx to localize to the mitochondria in the cell and whether this localization impacts the protein’s role in cellular replication.  The way in which we are testing this is by designing a series of primers to create mutant HBx forms that when amplified by PCR, will determine where in DNA.  Therefore, Joe, Kennadi and I all designed individual primers to determine what sequence of roughly 10 amino acids causes HBx to localize to the mitochondria.  We had to design both a forward and reverse primer for each reaction, that would "delete" about 10 amino acids from the HBx genome.  We used a program called SnapGene to make the forward and reverse primers necessary for mutating HBx.  Finally, we had Dr. Bouchard place our order of customly designed primers through a company called ThermoFisher.

My three forward primers

Day 3 was an exciting day for all members in the lab, as it was the last day for one of the grad students.  We had a mini celebration for her and ate Beiler’s doughnuts.  On the science side of things, I made agar plates and Lysogeny broth (LB broth) for later use.  Both were pretty simple recipes, although the agar plates did require an extra step since we had to make ampicillin for them.  After making them, the agar plates went in a freezer while the LB broth was left at room temperature on the lab benches’ shelves.  These two substances are to be used for growth of bacterium after we add our mutant protein’s to them.  Finally, Joe introduced us to the autoclave machine, a piece of equipment that heats up to extremely high temperatures, as we needed it to make our LB broth.

My LB broth on the shelf

Thursday was similar to Wednesday in the sense that I made substances to be used later as we await the arrival of our primers.  I made collagen plates, again for later use.  The 3x collagen will provide our mutant cells with something to adhere to as they grow.  After that I helped oe change the cell media growth for his Hepu2 cells he is growing for a separate project.  Since there wasn’t much else to do this day, Joe had Kennadi and I use the autoclave machine to clean all the dirty glassware in the laboratory.  When Dr. Bouchard saw this he joked that it was an abuse of child labor on Joe’s part.  Fortunately for me the sequence of shorter days in a row helped me readjust back to New Jersey time and my daily routine.  It also gave me a chance to go to the Reading Terminal Market on Thursday June 22, one of my favorite places in Philly.

Cleaned dishes coming out of the autoclave

The last day of the week was cut short for me since I had a dentist appointment.  And as murphy’s law would have it, this was the day that the primers arrived.  I spent all morning on Friday going through the lab safety training program that was required for Drexel research lab interns such as myself.  As I was preparing to leave, Joe gave Kennadi and I the news that our primers had arrived and that we’d be starting the first round of PCR that day.  I was able to pitch in a little before I had to leave my labmates to finish the rest of it.  That said I didn’t miss too much since I’ve done PCR a number of times before in Biotechnology.

Overall, it was a very smooth transition for me and I am really happy to be a part of this lab.  That said next week should have a lot in store for me as we can now progress with our project.