In this S10 proposal, we request funds for a QUANTSTUDIO 12K FLEX OPEN ARRAY REAL-TIME PCR SYSTEM manufactured by Thermo/Life Science/ABI that will enable BMC and Boston University users to carry out studies involving gene expression analysis, genotyping, microRNA profiling, and other RNA research projects. This innovative and versatile system, will replace the old and obsolete RT-PCR cyclers within the Analytical Instrumentation Core (AIC) that are no longer being supported by the manufacturer. Over the past eight years, more than 50 principal investigators and their laboratories have used the AIC's qPCR instruments. This new system will not only allow researchers to continue their on-going studies, but it will also create opportunities for new and original projects. In addition, having the latest software analysis tools and the additional capabilities of the new instrument will further promote the diversity and efficacy of nucleic acid research. It will allow the users to perform a wider-range of assays using different detection platforms at both the micro and nano- scales much more efficiently and will enhance data processing. Dr. Kenneth Walsh is the Principal Director and Principal Investigator (PD/PI) for this team project and is a leading scientist in the field of cardiovascular disease. His laboratory examines the molecular events that promote cardiac cell muscle growth, starting from differentiation and ending with cell death. Dr. Bryant will continue to use the AIC to determine the contribution of natural genetic variation on the mechanisms that confer susceptibility vs. resilience in substance abuse addictions. Dr. Myers will take advantage of the new qPCR system by conducting miRNA profiling to further examine the role of genes and gene regulation in both Huntington's (HD) and Parkinson's Diseases (PD). The above three scientists composed the Major User Group. The new instrument will enable other BU investigators (both major and minor users) to examine the molecular mechanisms involved in various areas of research that include: metabolic heart disease (Dr. Colucci), Alzheimer's Disease (AD) and Amyotrophic Lateral Sclerosis (ALS) (Dr. Wolozin and Dr. Qiu), diabetes (Dr. Ruderman and Dr. Rameh-Plant), ischemic limb vascularization (Dr. Matsui), obesity (Dr. Jing), calcium signaling (Dr. Bolotina), breast cancer and melanoma (Dr. Thiagalingam and Dr. Cui), dental disease (Dr. Gibson), sexually transmitted diseases (STDs) (Dr. Wetzler), virology (Dr. Connor) and hepatic disease (Dr. Bachschmid). The new instrument will also be used by AIC personnel to continue being able to provide the ?Gene Array Service? for all members of the BU research community. The new instrument will be housed within the AIC's qRT-PCR facility which is located on the 2nd floor at 670 Albany Street. The AIC has extensive experience when it comes to maintaining instruments and ensures that they are in excellent working condition with very little down time. Finally, the AIC has a long track record of having excellent equipment usage when it comes to RNA studies.
Quantitative qPCR is the gold standard when it comes to the quantification of gene expression and is the premiere method for validating microarrays or RNA-sequencing results. It is an immensely value tool that is applicable to virtually all areas of research and is especially useful for exploring pathways involved in cardiovascular disease, obesity, substance abuse disorders and even in neurodegeneration. As such, qPCR is vital in the discovery of novel biomarkers and in development of novel drugs.
