Assessment of Chemiluminescent Detection System for Forensic DNA Restriction Fragment Length Polymorphism Analysis
The purpose of this project was to determine whether a chemiluminescent detection system can give the sensitivity of 32P isotopic detection for human DNA restriction fragment length polymorphism (RFLP) analysis and have the speed, safety, and reduced cost of colorimetric detection. The assessment was made on the basis of: cost, sensitivity, time of operation, ease of operation, and ease of multiple probings. Human DNA was extracted from whole blood and processed for RFLP analysis according to the December 1990 FBI Protocol: "Procedures for the Detection of Restriction Fragment Length Polymorphisms in Human DNA." Both the "Boehringer Mannheim Genius" system which uses digoxigenin labeled probes detected with alkaline phosphatase-conjugated anti-digoxigenin antibody, and the "Promega Gene Print Light" system which uses alkaline phosphatase bound directly to the probe, were examined. Probes pH30, YNH24, and TBQ7 were used. Visualization was with "Lumi-Phos" 530 and Kodak XAR X-ray film. Results show that chemiluminescent detection is better than colorimetric or 32P detection in all aspects but sensitivity.
64 leaves. Advisor: Dr. Dean Hoganson