Research Administration & Mentoring Core (RAMC)
The Research Administration and Mentoring Core provides the leadership which ensures that scientific exchange, mentoring, and education thrive within bioMT. In addition to supporting and advising bioMT investigators, providing administrative and fiscal support, identifying strategic research opportunities, and fostering a strong scientific community, the RAMC will oversee a structured mentoring program, facilitate robust scientific exchange, and leverage institutional resources, which will enable bioMT to thrive as a nationally recognized COBRE.
Director: Dean Madden, Ph.D.
Contact: Cindy Stewart
Molecular Tools Core (MTC)
The Molecular Tools Core is a bioMT and Dartmouth community resource that enables research projects via acquisition or production of high quality recombinant proteins. We are developing optimization strategies spanning the molecular, cellular, and process levels in order to build an integrated pipeline capable of producing a wide spectrum of proteins and protein domain fragments.
PROTEIN PRODUCTIOn and characterization: from gene to crystal…
To provide the highest quality molecular tools for a specific project, the MTC will advise and work closely with researchers on construct design, expression host selection and on polishing strategies to produce large amount of pure proteins. The MTC will also execute or develop novel protocols for the production of recombinant proteins. Currently, we are producing proteins in mammalian cells and E. coli and will include insect cell and yeast expression systems in our toolbox in the near future. We are actively developing procedures to provide a robust quality control and characterization data package with the finished product, including spectroscopies and measurement of biomolecular interactions in close collaboration with the Molecular Interactions and Imaging Core (MIIC). The MTC recently incorporated the Crystallography Core Facility (CCF), providing the bioMT researchers the unique opportunity of designing, expressing, purifying, and crystalizing high quality proteins all in the same location.
The MTC provides specific training tutorials to facilitate technology transfer of developed processes and more generalized training to enable the community to independently use the available equipment.
AVAILABLE EQUIPMENT IN THE MTC:
(for a comprehensive list, visit RaDar)
•AKTA Pure 25 chromatography systems for purification of native and recombinant proteins equipped with:
• Multi-wavelength UV and fluorescence detectors.
• Capillary loops, superloops, and sample pumps for loading small and large volumes of sample.
• Pre-packed columns for size exclusion, ionic exchange and affinity chromatography.
•Beckman Avanti J-25and Optima L-70 centrifuges for low- and high-speed centrifugation.
•Bio-Rad Trans-Blot Turbo and Mini Trans-Blot Cell for membrane transfers.
•Bio-Rad PROTEAN IEF Cell for isoelectric focusing.
•Horizontal and vertical gel electrophoresis systems for protein and nucleic acid analysis (Bio-Rad and Invitrogen).
•Shaking incubators for cell culture (37ºC and low temperature).
•Electroporator for fungal, bacterial and eukaryotic cells.
•Microfluidizer LM10 for cell disruption.
Available equipment in the CCF:
(for more information visit RaDar)
•NT8 Drop Setter automated nanoliter/microliter drop dispensing.
•Formulator Screen Builder automated preparation of custom crystallography screens.
•Rock Imager images screen plates at scheduled intervals and allows users to remotely monitor drops.
Contact: Zdenek Svindrych, Ph.D.
We provide microscopy support to the bioMT community. With a broad range of microscopes available at Dartmouth (either located in one of the imaging cores – Class of 1978 Life Sciences Center and Dartmouth-Hitchcock Medical Center, or in individual labs), we can cover many applications. The MIIC uses microscopy for both biochemical and cellular studies, and will help individuals design and implement methodologies to answer specific research questions.
•Laser scanning confocal (Nikon A1RSi with FLIM/FCS, Zeiss LSM880 with Airyscan, Zeiss SLM800 with Airyscan).
•Spinning disk confocal (AndorDragonfly 302, AndorW1 with Mosaic and Micropointphotostimulation/photoablation).
•Total Internal Reflection (Nikon/AndorTIRF, Olympus TIRF).
•Mutliphoton (Olympus FVMPE-RS with MaiTaiDeepSeeand FLIM/FCS).
•Epifluorescence (Nikon Ti-E - inverted, Nikon 90i – upright, Nikon SMZ1500 – dissecting stereo zoom).
Available image-processing software:
•Amaris workstation with Measurement Pro, ImarisTrack, ImarisColoc, ImarisXT, ImarisCell, ImarisVantage, and Filament Tracker.
•NIS Elements workstation with Deconvolution.
Molecular Interactions and Imaging Core (MIIC)
INTERACTIONS AND IMAGING
To quantitatively characterize the molecular interaction of biomolecular systems, the MIIC will provide expertise in the sample preparation (working with the MTC), data collection, and analysis for a range of instrumentation including isothermal calorimetry, surface plasmon resonance, fluorescence-based measurements (fluorescence polarization, alpha-screen), and nuclear magnetic resonance. The MIIC has equipment available for bioMTmembers for label-free analysis of biomolecular interactions, including drug target identification and validation, and for numerous imaging applications to characterize biological systems.
Available equipment in the MIIC:
(for more information visit RaDar):
•Biacore X100 to measure biomolecular interactions in real-time.
•LI-COR Odyssey for quantitative, highly sensitive near-infrared fluorescence detection.
•Bio-Rad GelDocXR for high-resolution nucleic acid and protein gel imaging.
•Synergy Neo2 Hybrid Multi-Mode Microplate Reader for numerous HTS applications with multi-detection modes.
•DeNovixDS-11 FX+Spectrophotometer/Fluorometer for a variety of micro-volume nucleic acid and protein analyses.