Rae Ann Ciszewski 
Cardinal Stritch University
Milwaukee, WI
Faculty Mentor: Dr. James S. Maki

Yellowstone National Park is an area rich in microbial life and diversity. The objective of my project was to isolate and characterize thermophilicbacteria from Yellowstone National Park using molecular techniques. Thermophilic heterotrophic bacteria (growth at 55 degrees Celcius) were collected from a variety of hydrothermal vents in Yellowstone Lake and other thermal areas of Yellowstone National Park and cryogenically frozen (-20 degrees Celcius). Fifteen of thirty-three cultures displayed visible growth after rescue on Castenholz Trypticase Yeast Extract medium incubated at 55 degrees Celcius. Bacteria were grown and isolated in both liquid and solid forms of this medium. GELRITE was used as the gellingagent for the solid medium. After multiple streakings, the isolation of single colony types resulted (pure cultures), and DNA was extracted from the bacteria. With the DNA, two types of PCR amplifications were used. First, Random Amplified Polymorphic DNA (RAPD) analysis of these pure cultures has begun in order to detect genomic polymorphisms and determine the possible relatedness between the isolated microorganisms. Second, the 16S rRNA genes were amplified by the Polymerase Chain Reaction (PCR). These 16S rRNA genes will be sequenced in an attempt to identify the microorganisms.