PCR-BASED DNA TYPING SYSTEM

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We propose to develop a method of DNA typing using the polymerase chain reaction (PCR). We will develop a collection of PCR primers which will amplify specific VNTR regions of human DNA. we will optimize conditions 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. The result will be a rapid, non- isotopic technique that can be used in forensic and paternity analyses and frequency of alleles for different ethnic groups. This will allow determination of probability of exclusion and probability of identity in paternity, forensic and criminal studies. In phase I we will demonstrate the feasibility of this system by developing a pair of PCR primers for one of our highly polymorphic probes that contains the VNTR. We will further demonstrate our ability to create PCR- based tests by developing primers for a probe that does not contain the VNTR. In addition we will optimize conditions for electrophoretic separation of fragments resulting from amplification using primers from our model system. In phase II we will finish development of PCR primers for our entire set of paternity/forensic probes, develop a database for data analysis and institute the use of a video imaging system and software for data collection and storage.