The typing of PCR-amplified DNA by high-performance liquid chromatography

Human identification through DNA analysis has undergone tremendous changes since the first criminal conviction in the United States using DNA evidence in the Florida v. Andrews case of 1987. DNA is commonly used as forensic evidence to link suspects to crimes, exclude falsely accused suspects, reveal serial crimes, distinguish copycat crimes, identify the remains of a victim, and reconstruct accidents. In fact; DNA analysis is used in approximately 10,000 new criminal investigations annually. With the advent and use of the polymerase chain reaction (PCR) as a means to increase the copy number of specific DNA sequences, the use of DNA analysis as evidentiary material will continue to increase. Currently, gel electrophoresis is the DNA analysis method most commonly used. However, gel electrophoresis is a slow technique that typically takes more than two hours to complete, and once the electrophoresis is complete, the results can take days to process. In this dissertation, high performance liquid chromatography (BPLC) is used as a rapid DNA sizing/typing method. The chromatographic conditions for the separation of dsDNA were optimized. Using the optimal conditions for the separation of dsDNA, PCR products were sized using restriction enzyme fragments. In addition, a typing method for PCR products from the HUMTHO1 locus was accomplished. Finally, PCR products from different loci were multiplexed and typed using HPLC.