Determining the Primers and Methods for T-RFLP Analysis of Yellowstone National Park Soil and Soil from Washington and Lee's Campus to be Used with the LI-COR 4300

Abstract

At the onset of our research, our goals consisted of determining the primers that should be used in the T-RFLP protocol with the LI-COR 4300, optimizing the conditions for soil DNA extraction, PCR, and T-RFLP, and ultimately testing the methods on soil samples in order to obtain results concerning which populations of bacteria are present and in what quantity. The importance of perfecting the methodology used with the LICOR 4300 for Dr. Bill Hamilton's lab at Washington and Lee University is that it will provide a resource for comparing soil communities. Comparison of communities will provide information as to what species are present and ultimately how the communities change in response to surrounding environmental factors. The variations seen among different soil samples could provide more clear insight as to how the aboveground and belowground biotas interact. The comparisons may also help explain the exact impact that changes in nitrogen, carbon, oxygen, and mineral levels may have on the microbe community. The effects ofherbivory can thus be examined as well considering that herbivory incorporates numerous interactions that are often studied separately. [From Purpose]