Deepak Salem, May 12th, 2015
I decided to do my practicum on research – more specifically phytopathological microbiology. Over the summer of 2014, I started working at the United States Department of Agriculture (USDA) at the Sustainable Agriculture Systems Laboratory under Drs. Dilip Lakshman and Jaba Mukhopadhay (my mentor/site supervisor). I initially found the research position through a connection – my uncle is a researcher at the USDA as well working on food borne pathogens. I reached out to him and was able to land this research position at the USDA.
This was my first time doing non-descriptive research, so the first thing for me to do was simply shadow my mentor around the lab. I started to learn how to get work done around the lab and started to learn about the various microbiology and molecular biology that the lab focuses on. The first thing I learned how to do was plating. I learned how to prepare a plate appropriate for growing most fungus and bacteria using a Potato-dextrose gel, starting with preparing the gel, autoclaving it, then gently pouring it. Then, I learned how to transfer bacteria and fungus from on old plate to a new plate. That was the start for me in getting into the world of microbiology.
At this point, I had been mostly only viewing how my mentor Dr. Mukhopadhay (I called her Jaba) was conducting her molecular biology assays. This is when I started to really understand what was going on (after I had taken my first college genetics and cell biology classes). I learned how to operate a PCR (polymerase chain reaction) machine, operate a gel electrophoresis machine, properly centrifuge, apply blotting techniques (we dealt mostly with DNA/ RNA so mostly Southern/Northern Blotting), apply Ethidium Bromide staining to a gel for analysis, utilize genome analysis software, and most importantly how to clean up everything!
After learning all of the basic techniques (and the complicated ones), I started actively working on various ongoing projects in the lab. At this point, Jaba was actively working on the completion of a cDNA library (a method of keeping all of the genetically modified versions of the bacteria/fungus that we used in an easy to access place) so I took an active role in completing it as well. Completion of a cDNA library requires a lot of DNA analysis, so I thoroughly learned how to prepare DNA samples for analysis and quantification via kit or by scratch (mini-scale and macro-scale). Then, I would plate all the bacteria/fungus that had good matches for their DNA and place them in our library.
I started work on my first major experiment in spring of 2015. We call it the “Biocontrol Bacteria” experiment. The lab’s main focus is on diseases of the soybean and the plant roots, which is heavily affected by a bacteria known as Rhizoctonia solani. We labeled and infected a plot of land that had soybeans growing in it with R. solani. We noticed that the plot of land still had soybeans growing, which is unusual. So we analyzed to see whether the R. solani was potent, which it was. Afterwards, we deduced there must be another bacteria that is in that plot of land that is killing the R. solani. So we spent about 3 months conducting isolation experiments to determine which of about the 20 million bacteria in that plot of land was killing it. The experiment is still ongoing and about the concluded and sent to publishing.
I learned a lot about research. The most valuable thing from this internship is the fact that I have learned practically all of the basic laboratory techniques of any lab. Essentially, it prepared me to be able to work in any lab ever. It also made me more aware of how people view things like GMOs and such. GMOs, according to my knowledge, are beings that I work with every day and they are not necessarily bad things. I also learned extensively about how the process of research and how the process to paper is like. I really also learned how intensively long the process is as well. Working in this lab has given me much appreciation for research and working in a lab, but it also alerted me to the lifestyle of working in a lab. It solidified my initial plans of proceeding to medical school at the end of my college career.