| Twelve strains of Acidifhiobacillus ferrooxidans were recovered from acid mine drainage (AMD) sites from three different geographical locations: Copper Cliff, Ontario, Canada; Minerai City, Ohio, USA; and Cornwall, England. The spread-plate technique and various solidified media were used to isolate and purify all strains. Once purified, each strain was grown in TK liquid medium (Tuovinen and Kelly, 1973). After reaching stationary phase, DNA was extracted from each purified culture and DNA concentration was estimated using spectrophotometry. Twenty 10-mer primers from Operon Technologies Inc. were used for the polymerase chain reaction (PCR). Amplification reactions were done using 7 ng of DNA in a Perkin-Elmer DNA Thermal Cycler. The amplification products were separated by electrophoresis on 1.5% agarose gels stained with ethidium bromide, and photographed under UV light. The RAPD (Randomly Amplified Polymorphic DNA) profiles were compared on the basis of the presence or absence of each DNA band and a data matrix was constructed. (Abstract shortened by UMI.)... |
