Jerry Wang, Week 3, PCR, PCR, PCR and more PCR

Week3:

                This week was really exciting since my primer finally showed up at the doorstep right when I entered the lab on Monday. The first step of processing such primer is to reconstitute it. I was really confused by the word “reconstitution” since I have never heard of such word in my entire life. This is actually a common experience since research labs often use very advanced word for very simple idea. And in this case, reconstituting a primer literally means adding water to dissolve the DNA piece that was laying at the bottom of the test tube. I “reconstitute” the primer in the fancy fume hood we got in tissue culture lab. A fume hood is basically a sterile environment that would prevent particles from entering or exiting the hood. Here, since the primer is very delicate and we would not want any contamination to it, we performed the reconstitution process with molecular biology grade water (just extremely pure water) inside the fume hood.

                After that, there is the PCR process. After PCR, I ran a gel electrophoresis to see if my primer worked or not. This is the most nervous moment across the entire summer, not only because none of my AP Biology PCR attempt back in sophomore winter had worked, but also because if the primer did not work (even though in principle it does not have any reason to fail) I would have to redesign the primer which would then arrive next week. If I had learned anything so far from EXP, it is that time is the most valuable thing in the lab. Thankfully, my primer turned out alright. As you can see in the picture, besides the primer dimer at the bottom of the gel, my primer amplified three genetic sequences. CD3 epsilon’s mRNA sequence is around 600 base pairs. So, I cut out that part of the gel and performed gel extraction in order to extract the CD3 epsilon’s encoding DNA from the gel. Sadly, this was my first time ever to perform such operation and there were some minor mistakes that greatly impacted my final result. In the end, my final product was highly contaminated with low DNA concentration. Therefore, I need to redo the entire process again.

                On Friday, our entire lab walked to the human translational research center to attend a joint lab meeting. In principle, what we are doing in our lab is very similar to our human counterpart. It’s just that canine patients and cancer research get a lot less attention compare to human patients, which is very understandable, so we receive much less grants compare to the human translational research lab. Their building was fascinating: big working area, separate lab area, gigantic meeting room, well supplied coffee machine, everything in that building feels so professional and high-end. The joint lab meeting took place on the floor where Dr. Carl June invented CAR T cell. For some unknown reason, this small trip to the human translational research center felt like a “pilgrimage” to me, since it was there where the miracle happened, where CAR T cell was first invented.