High-Throughput Protein Lab
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 titering recombinant baculovirus.
Protein Purification Facility
We have facilities to purify 1–100 milligrams of 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 fast protein liquid chromatography machines — an Acta Purifier-10 FPLC and an Academic FPLC from Pharmacia — and two high-throughput AKTA xpress units equipped with a variety of ion exchange, gel filtration and affinity columns.
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 nanoliter drops are available for rapid screening of hundreds of conditions using small quantities of macromolecule. One unit is dedicated to protein crystallization; one to RNA and RNA-protein complexes and one is equipped for LCP crystallization.
Our X-ray crystallography facility includes an X-ray diffraction system: Rigaku RU-300 copper rotating anode equipped with Raxis IV dual imaging plate detector, High Flux confocal optics (Molecular Structures Corp.) and an X-Stream cryo-cooling system, as well as Linux computers for data collection, modeling and structure determination.
LS-CAT Synchrotron Beamlines
In addition to the on-site facilities, U-M 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. Each station is equipped with state-of-the-art diffractometers, robotic sample changers, and large-area CCD detectors.
- The primary station, 21-ID-D, is fully MAD capable and is tunable from 5–20 keV. (Equipped with an Eiger 9M detector.)
- 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.
Beam diameters of 5 to 50 microns are available. Additionally, mail-in and remote access services are available.