Principal Investigator/Program Director (Last, First, Middle):
RESOURCES
FACILITIES: Specify the facilities to be used for the conduct of the proposed research. Indicate the performance sites and describe capacities,
pertinent capabilities, relative proximity, and extent of availability to the project. Under “Other,” identify support services such as machine shop,
electronics shop, and specify the extent to which they will be available to the project. Use continuation pages if necessary.
Laboratory: Located on the third floor of the UM Life Sciences Institute (LSI), the 2,260 square-foot UM
Center for Structural Biology provides expertise and specialized laboratories to researchers who wish to
produce biological macromolecules or determine their crystal structures.
High-Throughput Cloning and Expression Testing: A high-throughput cloning and protein expression
facility aids researchers in identifying protein constructs that are stable, soluble and suited for structural
study or for assay development. The HTP Lab is equipped with a Beckman Biomek dual-arm liquid-handling
robot, a Caliper Labchip 90 electrophoresis instrument and ancillary equipment for performing DNA and
protein manipulations, bacterial culture in 96-well plate format and baculovirus-insect cell infection in 24-well
block format. The facility integrates semi-automated PCR, cloning, transformation, colony picking, protein
expression, isolation and quantification. A Roche MagNAPure and Lightcycler QPCR instrument are used
for tittering recombinant baculovirus.
Protein Production: The CSB provides facilities to purify 1 – 100 mg protein. In the facility, there is a
fermentation room with multiple variable temperature shakers for large- and small-scale fermentation, and a
French press, sonicator and microfluidizer for breaking cells. The large wet lab has two FPLCs (Acta Purifier10 FPLC and an Academic FPLC from Pharmacia) and 2 high-throughput ÄKTAxpress units equipped with a
variety of ion exchange, gel filtration and affinity columns.
X-ray Crystallography Facility: For crystallization, two environmental rooms adjacent to the wet lab are
used for crystal growth at 4 °C and 20 °C. Each room is equipped with stereomicroscopes for crystal
viewing. There are two Gryphon crystallization robots capable of dispensing 100 nL drops are available for
rapid screening of hundreds of conditions using small quantities of macromolecule. One unit is dedicated to
protein crystallization; the other to RNA and RNA-protein complexes. The on-campus X-ray crystallography
facility includes two X-ray diffraction systems: two Rigaku RU-300 copper rotating anodes equipped with
Raxis IV dual imaging plate detectors with High Flux confocal optics (Molecular Structures Corp.) and XStream cryo-cooling systems, and Linux computers for data collection, modeling and structure determination.
LS-CAT Synchrotron Beamlines: In addition to the on-site facilities, UM is a member of LS-CAT, a
consortium of academic and research institutions that operates a multi-beamline facility located at Sector 21
of Argonne National Laboratory's Advanced Photon Source. This facility consists of four experimental
stations for X-ray crystallography with insertion device sources. The primary station, 21-ID-D, is fully MAD
capable and is tunable from 5–20 keV. The 21-ID-F and 21-ID-G stations are at fixed energy (12.67 keV)
and are suitable for selenium SAD experiments. The fourth station, 21-ID-E, is planned to be tunable
for selenium and bromine MAD. Each station is equipped with state-of-the-art diffractometers, robotic
sample changers, and large-area CCD detectors. Beam diameters of 5 to 50 microns are available.
Additionally, mail-in and remote access services are available.
Clinical:
Animal:
Computer:
3 GHz Dell linux box, 2 GHz IMAC running OSX
Other:
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February, 2014
Principal Investigator/Program Director (Last, First, Middle):
MAJOR EQUIPMENT: List the most important equipment items already available for this project, noting the
location and pertinent capabilities of each.
Two Rigaku RU-300 copper rotating anode equipped with Raxis IV dual imaging plate detector from
Molecular Structure Corporation with accompanying Linux computer for data collection and processing
Two X-Stream cryo-cooling system
Akta Purifier-10 FPLC from Pharmacia
Akta Academic from Pharmacia
Two ÄKTAxpress units from GE Healthcare
Continuous French Press
Sorvall Superspeed Centrifuge with rotors
Beckman XL-1 Analytical Centrifuge
Optima L-90K Beckman Coulter Ultracentrifuge
Protein Solutions Dynapro Dynamic Light Scatter
Two Gryphon Crystallization Robots
Beckman Biomek FX
Caliper LabChip 90 for PAGE analysis of protein and DNA samples.
Octet Red using Bio-Layer Interferometry technology
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February, 2014