Shivani Gupta- Week 2- Epigenetics- Reddy Lab

                                 This week, I started to plan for cloning using a program, pDRAW32, that lets you clone DNA fragments virtually by using restriction enzymes. We are cloning three different protein coding genes, YY1, Lap2B, and lmn A. While it took us a couple of hours to determine how we can clone these genes, it was exciting learning and becoming more comfortable with this software.  This week, I also did some tissue culturing, which I became much more comfortable doing under the hood with sterile technique.

On Wednesday, one of the students in my lab, Lauren, had to give a presentation for the internship program she is part of. At the seminar, I learned more about her project, which is closely connected to mine. At the seminar, a classmate of Lauren’s also gave a presentation about the connection between depression, platelet serotonin, and coronary heart disease. On Thursday, Jen, my mentor in the lab, did a western blot which consists of gel electrophoresis to locate the antibodies from which the proteins have been separated. On Friday, I had the chance of doing a Western blot by myself. On Friday, we also transfected cells and completed a Western on the DNA. After the gel ran through, we had to cut the gel under UV light and then had to melt the gel. We ended up waiting for an hour before we realized that our gel wasn’t melting because the heater was broken! After we finally melted the gel, we did gel extraction which consisted of a series of washes and running the samples, YY1, Lap2B, and lmnA, on the centrifuge. On Friday, I also went to Happy Hour, which was a chance to network with people in other research departments at Johns Hopkins. I got to meet one of the grad student’s former PI and a grad student who is also working in the epigenetics department under Dr. Taverna.

This week went by so quickly and I am looking forward to the coming weeks at my lab.