Equipment Requested: This proposal seeks support for acquisition of a Pyromark Q96 MD automated pyrosequencing system made by Biotage/Qiagen. The system will allow high-throughput quantitation of DNA methylation in epigenetic studies by a large number of NIH-supported researchers. Rationale: Pyrosequencing is a real-time 'sequencing-by-synthesis'technology that is especially useful for quantitative analysis of DNA methylation. The user group conducts studies of environmental agents and the mechanisms of their health effects, including comparative analysis of epigenetic modifications linked to developmental, neoplastic and functional changes. Pyrosequencing will offer the investigators the ability to perform high-throughput and quantitative analyses of global and site-specific methylation with great precision. Implementation: The instrumentation will be situated within a well-established core facility in current operation and supported by a NIEHS Center grant. The instrumentation will be managed by the PI who directs the NIEHS core facilities, and has extensive experience in core facility operation for user benefit. Two senior technologists with experience in cell and molecular biology will be trained on the system and will provide training, supervision and support to the user group lab members. Support includes space for the instrumentation within the core facility (institutional), coverage of service contracts and future component replacements for the system (institutional), and core facility staff support (NIEHS Center grant). An internal advisory committee will supervise use and assure access to the system. Summary: This instrumentation will significantly improve the research capacity and productivity of a large number of NIH-supported investigators via an institutionally supported, well-established core facility.
