We are requesting a Roche real-time polymerase chain reaction (PCR) Workflow System with automated components for preparation of tissues (MagNA Lyser), isolation of nucleic acids (MagNA Pure Compact), and a real-time PCR (LightCycler 480). The LightCycler 480 is a high throughput real- time PCR instrument that detects and quantifies nucleic acid sequences, and has a 96- and 384-well plate capacity. This PCR Workflow System is very easy to use, is accompanied by powerful software, and enables an investigator to go from tissue to nucleic acid quantitation in 3-4 hours. This instrument will be housed in the MicroArray Core Research Facility of the University in the Bosomworth Health Sciences Research Building, and will be used by faculty in the Graduate Center for Toxicology, and in the Department of Anatomy and Neurobiology. The group of investigators participating in this application includes seven faculty with NIH-supported grant projects who are major users, and three additional faculty who are minor users. The research projects are diverse, but revolve around three major themes: 1) oxidant-stress induced tissue injury and cancer, 2) DNA damage, repair and mutagenesis and 3) hormonal regulation of gene expression. We intend to use this instrument for the high-throughput real-time quantitative analysis of RNA transcripts and DNA targets in biological samples and in detection of single nucleotide polymorphisms (SNP) in genes important in cancer. As indicated in the research projects from each investigator, the availability of the proposed instrument will significantly improve the scope and timeliness of their studies. Most importantly, it will allow the science to determine the pursuit of exciting discoveries in their funded research projects, rather than availability of current instruments. The overall goals of this shared multidisciplinary research facility are to use this advanced technology resource to carry out cutting-edge research that characterizes today's biomedical research and to train our doctoral students and postdoctoral fellows in the use of these modern methods that will enable them to develop into successful independent investigators. The research problems addressed by our investigators are highly relevant to major public health problems related to cancer, both in the mechanisms of tumor formation, the reasons for (lack of) effectiveness of cancer chemotherapeutic drugs, and the regulation of genes by hormones that are important in women's health. ? ? ?
