Last week I explained why I am doing the research that I am doing, and the significance of such research. In week 3 I promised to provide some details on the procedures involved in this research, so this week I will explain a little about polymerase chain reaction (PCR). PCR is the process used to amplify fragments of DNA that have been extracted from a specimen in order to make millions of copies to prepare for sequencing. To do this, we choose a particular gene or fragment of a gene to target. In this case, it is the gene that encodes a large subunit ribosomal RNA, 16S. This gene region is adjacent to highly conserved regions of DNA, and therefore we can use “universal” primers to start the process of transcription (Palumbi et al., 1991). Besides primers, we also need the nucleotide building blocks of DNA, DNA polymerase that can withstand high temperatures, and of course, a template strand of DNA that we want to copy. Once these are all mixed in the correct quantities, we can begin the process of PCR. Amplification happens through repeated cycles of heating and cooling; heating to 95ºC in order to denature or separate the two strands of DNA in the template, followed by cooling to 50ºC to allow the primers to anneal to the separated strands of DNA, then warming to 72ºC to allow the DNA polymerase to elongate the DNA. This process is repeated between 25 and 40 times, producing up to a million copies of the DNA (Rapley, 1998).
This week officially marks the halfway point of the OIMB REU program for 2017; where has the time gone? With each day that passes, I am increasingly thankful to have this opportunity to gain valuable research experience that will greatly benefit me as I move forward into a science-based career. This program has given me the confidence that I needed to finalize my degree path, and because of this program, I have decided to pursue a minor in biology in addition to my bachelor’s degree in natural resources. I am looking forward to what the second half will bring.
This past Friday, the REU interns were treated to a tour of the biology department at the main campus of UO in Eugene, where we were allowed to observe multiple studies across a variety of labs. While on campus, we also spent time exploring the UO Museum of Natural and Cultural History where we observed features of geological processes across the state of Oregon, artifacts used by early people in the area, as well as some thought-provoking photographic exhibits; I would recommend this stop to anyone going to the Eugene area.
Palumbi S, Martin A, Romano S, McMillan WO, Stice L, Grabowski G. (1991) The simple fools guide to PCR Version 2.0. Honolulu, HI: Department of Zoology and Kewalo Marine Laboratory, University of Hawaii.
Rapley, R., & Walker, John M. (1998). Molecular biomethods handbook. Totowa, N.J.: Humana Press.
My name is Becky, and I have been local to the Coos Bay and Charleston area for two years with my boyfriend David, and my dog Mojo. I moved to this area to complete my associate's degree at Southwestern Oregon Community College in anticipation of transferring to a four-year university. I now attend online at Oregon State University and I am planning a move to Bend, OR in August to continue at the OSU Cascades campus in order to finish my bachelor's degree in either natural resources or biology.