Practicum Reaction

Practicum Poster

May 11, 2010

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My project for the Practicum was the construction of a West Nile Infectious Clone. I worked within Dr. Brenda Fredericksen's lab located in 3126 Biosciences Research Building. I worked under the direct supervision of two graduate students, Lisa Injaian and Kat Pankow. My project site works with the West Nile Virus; the main focus of the lab is to identify and define viral factors responsible for increased pathogenesis among certain strains of West Nile Virus. They work with three strains in particular, New York, Madagascar, and Kujan. My project was to construct a chimeric infectious clone combining portions of highly pathogenic strain of New York 2000, and nonpathogenic strain, Madagascar 1978. An infectious clone is dsDNA copy of a viral genome inserted into a bacterial plasmid. This allows researchers to introduce site-specific mutations within the viral genome or construct chimeric genomes from two separate virus strains. The clones can then be used to generate recombinant viruses which can be tested in future experiments to determine their pathogenesis.

I really enjoyed working at this site. I had actually been working in this lab as a work-study student. However, I was just washing dishes, autoclaving breakers, and maintaining the lab. I did not actually get to do any science. However, this practicum gave me an opportunity to ask my professor for research experience. She approved and since everyone already knew me, the transition into research was pretty smooth. Everyone within the lab was very patient with me and would slowly teach me the methods of cloning. Dr. Fredericksen didn’t actually teach me anything; the graduate students, Lisa Injaian and Kat Pankow, taught me everything I know. Although my site supervisor didn’t teach me the techniques of cloning, she helped me to fully understand the project. At first, I was just doing what I was told and followed directions; I did not truly understand the process of what I was doing. My site supervisor sat down with me and went over all the little details. After talking to her, I finally understood what an infectious clone was, what the nature of my project was, and how to go about constructing the clone.

I learned a lot from this practicum in terms of scientific knowledge and application. This experience has taught me a lot about genetic engineering techniques and methods of cloning. It helped me apply the knowledge I learned from BSCI105 to real life application. Using techniques such as disgestion, ligation, PCR, and transformation and knowledge of bacteria vectors, I was able to help construct the infectious clone. I used restriction enzymes to cut my DNA, used Mini-Prep Kits to isolate my DNA, ran agarose gel electrophoresis to separate portions of my DNA, and used PCR and Gel Clean-Up kits to purify my insert portion of the DNA.

Through this experience, I have come to learn that failure was often associated with science. My experiments, mainly digestions and transformation, would fail many times. The bacteria would not accept my DNA, the vector and insert would not ligate together, the restriction enzymes would not cut correctly; there were a lot of thing that could go wrong. I am somewhat of a perfectionist so I could not accept this failure. I eventually became very stress, disappointed in my failure, and even thought about giving up. However, my lab reassured me that everything was ok. After many hours of work, I finally realize that by becoming stress by my failure, it would only hinder me further. I need to accept my failure and keep trying. Eventually my project worked it out and I got results. The second thing I learned was that science takes a lot of time. My cloning project took about two months and yet I was only able to move one step forward in my construction. I didn’t even finish my clone. Lisa pointed out to me that it took her a year once to construct one of her clones.

Beyond science, I’ve come to realize that actual experience is a necessity. Although I had done well in my biology classes, I realized that learning something in class is totally different from actually understanding it. I “learned” and memorized facts about genetic engineering and cloning from class, but I couldn’t apply it to my research. I needed Lisa and Kat to re-teach me and guide me. Although someone might think that he has learned something in class, he won’t truly understand it unless he has applied it to real life. More importantly, I realized that I would not like to become a scientist. As a child, I always wanted to become a scientist (I actually wanted to become a mad scientist and take over the world). Although this was an enjoyable experience, I cannot see myself doing this type of work for the rest of my life. I would be able to work like this for maybe a year or two, but not for life. My goal is still set to become a doctor.

I just want to take another moment to thank scholars for giving me the opportunity to gain this research experience. Through this experience, my supervisor has actually asked me to stay within the lab. I am currently working there as an undergraduate researcher and will continue to do research there this summer.

Last modified: May 11, 2010