E. coli bacteriophages were isolated from local Mount Vernon ponds and creeks. The phage strains were purified using a repeated double agar plating method. The pure phage strains were had a titer range of 5.0×10 8 to 2.0×10 11 viruses per milliliter observed. The viruses were then concentrated by ultra-centrifugation. DNA was isolated from the viruses by phenol:chloroform extraction. The viral DNA was cut with restriction enzymes and agarose gel electrophoresis was performed to observe restriction enzyme fragment banding patterns. A total of three different identifiable fragment banding patterns were observed. The three different strains were then used to begin development of polymerase chain reaction (PCR) and amplified fragment length polymorphism (AFLP) analysis protocols for bacteriophage DNA fingerprinting.
Donald Harker, ’04 Mt. Vernon , IA
Majors: Biochemistry & Molecular Biology
Sponsor: Jeff Cardon