A PCR-BASED DNA TYPING SYSTEM

A METHOD OF DNA TYPING USING THE POLYMERASE CHAIN REACTION (PCR) WILL BE DEVELOPED, AS WELL AS A COLLECTION OF PCR PRIMERS THAT WILL AMPLIFY SPECIFIC VNTR REGIONS OF HUMAN DNA. CONDITIONS WILL BE OPTIMIZED TO ENABLE VISUALIZATION OF THE RESULTING DNA FRAGMENTS ON A POLYACRYLAMIDE GEL. BY USING SEVERAL PAIRS OF PCR PRIMERS, AN UNAMBIGUOUS BANDING PATTERN WILL BE GENERATED FOR EACH INDIVIDUAL. IT IS ALSO PLANNED TO CREATE A DATABASE OF INFORMATION ON THE FREQUENCYOF ALLELES FOR DIFFERENT ETHNIC GROUPS. PHASE I RESEARCH WILL DEMONSTRATE THE FEASIBILITY OF THIS SYSTEM BY DEVELOPING A PAIR OF PCR PRIMERS FOR A HIGHLY POLYMORPHIC PROBE THAT CONTAINS THE VNTR. THE ABILITY TO CREATE PCR-BASED TESTS WILL BE FURTHER DEMONSTRATED BY DEVELOPING PRIMERS FOR A PROBE THAT DOES NOT CONTAIN THE VNTR. IN ADDITION, CONDITIONS WILL BE OPTIMIZED FOR ELECTROPHORETIC SEPARATION OF FRAGMENTS RESULTING FROM AMPLIFICATION USING PRIMERS FROM THE MODEL SYSTEM. IN PHASE II, THE DEVELOPMENT OF PCR PRIMERS FOR THE ENTIRE SET OF PATERNITY/FORENSIC PROBES WILL BE FINISHED, A DATABASE FOR DATA ANALYSIS WILL BE DEVELOPED, AND THE USE OF A VIDEO IMAGING SYSTEM AND SOFTWARE FOR DATA COLLECTION AND STORAGE WILL BE INSTITUTED.