The Molecular Biology Core Facility is a full-service, CLIA-certified genomics laboratory located in the James G. Cannon Research Center at Carolinas Medical Center (CMC), part of Carolinas HealthCare System. The Facility participates in and contributes to system-wide and regional research by performing various molecular biology techniques applied to different areas of health-related research. Projects are conducted as part of independent faculty research as well as resident research done as a component of the residency training program.
The Facility offers various levels of service, including performing for or consulting with individual investigators to implement standard experimental design and sample preparation protocols, as well as assistance with post-run data analysis and interpretation. The Facility has been serving the Charlotte Genomic Consortium in collaboration with UNC-Charlotte faculty, and has been the central location for a clinical study-related repository for various departments within our organization.
Mission Statement: To contribute to the enhancement of scientific research at Carolinas HealthCare System and the Greater Charlotte region as a whole, and to assist investigators with their acquisition of research results and funding by the introduction and implementation of state-of-the-art techniques to enhance their research capability.
Molecular Biology Facility
The Molecular Biology Core Facility is equipped with multiple Next Generation Sequencing instruments, sequencing library robotics, template preparation equipment, capillary sequencing instruments, two microarray systems, automated nucleic extraction instruments, a pipetting robotic system, a multiplex suspension array system, real time and conventional PCR instruments, low volume spectrophotometers, a CCD camera enhanced imager, and a vacuum speed concentration instrument.
The Facility has a state-of-the-art powerful computer workstation and a library of 3rd party data bioinformatics tools for the analysis of genomic data. The core also has a license for software for quality control and differential expression analysis of real-time PCR data.
Next Generation Sequencing
The Molecular Biology Core has Next Generation Sequencing (NGS) Instruments. NGS parallelizes the sequencing process thereby producing millions of sequences at once. With this technology, we gain the ability to globally investigate disease mechanisms including DNA sequence variants, RNA expression levels, and promoter methylation status on a high resolution sequencing platform. High-throughput sequencing has been employed to identify unknown causative mutations in human disease and will likely impact our understanding of complex disease trait loci and pharmacogenomics. The improved detection of rare sequence variants by high-throughput sequencing can also be applied to the discovery of novel somatic mutations in cancer. Such studies are expected to lead to a better understanding of the molecular pathogenesis of cancer, and will undoubtedly result in novel diagnostic and therapeutic approaches. Epigenetic regulation of gene expression underlies the pathogenesis of many diseases, especially cancer. NGS has been employed to detect and characterize DNA-protein interactions resulting in chromatin remodeling. Global analysis of RNA expression can enhance our understanding of both normal cellular physiology and disease states. Aberrant expression of small non-coding RNAs such as microRNAs has been implicated in the pathogenesis of disease as well. An advantage of the NGS approach for transcriptome analysis is that it is able to characterize all transcriptional activity, coding and noncoding, without prior assumptions thereby permitting the discovery of novel tissue-specific isoforms and the identification of previously undetected species. The Ion Torrent and Ion Proton instruments are scalable and flexible. The scalability is derived from the fact that the sequencing chips are essentially the sequencing machine. Thus, these instruments give the user access to several levels of throughput. Low-cost reagents and consumables coupled with customizable throughput levels result in a very low cost of operation compared with other NGS systems.
The Microarray Facility located in the Molecular Biology Core Facility was established by the Charlotte Genomics Consortium to serve investigators at both UNC-Charlotte (UNCC) and the James G. Cannon Research Center at CMC. The Facility assists individual investigators to design, implement, analyze and validate studies aimed at examining gene expression levels as well as performing whole genome analysis. Microarray data analysis software is also available for use by the investigators. Having been certified in the processing of expression analysis (including both 3' expression and exon) and DNA analysis (including high density SNP) arrays, multiple applications and fields of studies are supported.
Other Research Activities
The Facility has been serving as a tissue bank for various clinical departments. The specimen repository lab is equipped with a sample tracking system, BSI-II software which provides James G. Cannon Research Center and CMC investigators with data collection and specimen inventory capabilities.
Nury Steuerwald, PhD, HCLD (ABB), Director
Judy Parsons, Research Manager
Alicia Hamilton, Research Analyst
Amanda Schaller, Research Technician
Hiral Amin, Research Technician
Pooja Pardhanani, Research Technician
Nury Steuerwald, PhD, HCLD (ABB)