The X-ray crystallography Core provides a state-of-the-art facility for the generation of crystals and structure determination of macromolecules including proteins, DNA, RNA, and complexes between macromolecules and their ligands.

Sample Analysis:

Consultation and in silico analysis
1. Protein expression and purification
2. Domain structure, including disorder and secondary structure prediction

Physical Analysis
1. Native PAGE using GE HealthSciences PhastGel.
2. Limited Proteolysis and characterization by SDS-PAGE
3. Size Exclusion Chromatography
4. Sedimentation Equilibrium analysis by AUC
5. Sedimentation Velocity analysis by AUC.

Crystallization:
1. Set up crystallization trials at 4 ºC and/or 20 ºC.
     a. Initial trials (e.g., appropriate concentrations)
     b. Full scale trials (4 different 96 well factorials at 3 protein concentrations and 2
         temperatures)
2. Optimization – additive screens and factorial overlays
3. Automated Screening at 2 temperatures (scan daily for first week, weekly thereafter, finish experiment at 3 months)

Diffraction quality:
1. Test diffraction using capillary mounted crystal
2. Test/screen cryo-conditions

Data collection:
1. Collect, reduce and merge data.  Generate table of statistics
2. MAD/SAD phasing – help design, collect, reduce and merge MAD data
3. Generate table of statistics including anomalous dispersion differences

Structure Determination:
1. Solve structure by Molecular Replacement
2. Solve structure by MAD/SAD phasing
3. Refine structure
4. Produce relevant statistics (e.g., R and Rfree, RMS deviations)
5. Structural analysis (superpositions, electrostatics, etc)
6. Deposit Structure at PDB