(Investigator?s Abstract): Real-Time PCR has resulted in the ability to accurately and quantitatively measure nucleic acid with great sensitivity. This can be accomplished by 1) using TAQMAN technology which, using the exonuclease activity of TAQ enzyme, allows for the expression and measurement of a fluorescent signal that can be detected at each cycle of PCR amplification and 2) Molecular Beacon technology, a technique in which unfolding of a fluorescent molecule and quencher after binding to a complementary amplicon, also generates and measures a fluorescent signal during each PCR amplification. The ABI PRISM 7700 molecular sequence detector with a TAQMAN upgrade package has the capability of using these technologies. It has the capability of measuring 4 separate fluorescent signals simultaneously in the same reaction. This instrument has also been used to detect genetic polymorphisms using appropriately designed molecular beacons. Various investigators throughout NYU are already familiar with the power of the instrument and expect that access to the instrument will improve their ability to perform measurements crucial to their current or expected research. The instrument will be a shared resource utilized by individual scientists throughout the medical school. There is no comparable instrument within the entire medical school at present. It will be also utilized by the NYU Center for AIDS Research (CFAR) Core Virology Lab (directed by Dr. Borkowsky) which serves as a host of investigators engaged in AIDS Research and the NYU General Clinical Research Center (GCRC) which serves the NYU community of investigators engaged in clinical research. Usage of the instrument will be monitored by an internal advisory group representing the CFAR and GCRC. The cost of running assays will be minimized by charging only for reagent cost and instrument maintenance and will be subsidized by funds from the GCRC and from individual grants. The principal investigator will be responsible for helping design primers and probes for the instrument as well as insure that the assay and those who manage it undergo appropriate quality control procedures.
