SCIENTIFIC CORES
Bacterial Protein Expression Core
Biochemistry Core Cell Biology and Imaging Core Computation Core EM Crystallography Core EM Tomography Core Eukaryotic Protein Expression Core Fluorescence Spectroscopy Core Protein Interactions Core Protein NMR Spectroscopy Core RNA Structure and Dynamics Core Tissue EM Core Virology Core Virus Imaging Core X-ray Crystallography Core |
X-Ray Crystallography Core
Director: Chris Hill, PhD Managers: Frank Whitby, PhD and Heidi Schubert, PhD The X-ray Crystallography Core determines high-resolution structures of proteins and complexes. The Core is well equipped for all aspects of protein crystallography, and resources include a crystallization robot, crystal imagers, X-ray generator, detector, computers, and constant temperature rooms (4, 13, 20°C) dedicated for crystallization. We have excellent access to synchrotron radiation facilities and have material for shipping multiple sample-containing dewars to synchrotrons for remote data collection. We typically collect data via internet access within a few days of shipping crystals to the Stanford (SSRL) or Brookhaven (NSLS) synchrotrons, with dewars of crystals shipped every three to four weeks. Instrumentation and Capabilities Robot The X-ray Crystallography Core is equipped with a Robbins Scientific Phoenix crystallization robot. This specialized liquid handling instrument is ideally suited for efficiently setting up large numbers of 96-well crystallization trays. The robot distributes reservoir solutions from standard 96-well blocks that are either purchased commercially or made by lab members to sample desired crystallization conditions. Imagers We have two desktop Minstrel HT crystal incubation/imagers, one each in a dedicated 20oC and 4oC crystallization room. Bar-coded crystal trays typically reside in the imagers for 8 weeks, during which time drops are imaged at 12, 24, and 48 hours and then at 7 day intervals thereafter. We have space for 160 trays in each imager, so trays are removed after three weeks to make space for new crystallization trials. Generator/Detector Our in-house X-ray diffraction facility includes a Rigaku MicroMax-007HF generator with VariMax-HR confocal optic and a Rigaku R-AXIS IV++ image plate detector. The detector sits on a sled with maximum sample-to-detector distance of 300mm. Users are trained to collect data independently and instrument time is tracked. Data tracked for the last 20 months shows that the instrumentation has been running and available for data collection on average 89.4% of the time. Down time comprises routine maintenance and periodic repairs of the generator, detector, or cryostream, and changeover time between users. Computers We use primarily LINUX-based computer systems, either RedHat LINUX for PCs or Power-PC-based Macintosh computers which use a form of the UNIX operating system. In addition to the numerous Macintosh and PC computers of individuals in the lab, we have 6 PCs dedicated to crystallographic refinement, modeling, and other analyses in a common computer room along with numerous NMR-core computing resources. This room serves as a hub of crystallographic and NMR analyses where lab members interact and benefit from each other’s experience. We also have free access to the University of Utah Supercomputing Center on campus.
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