A procedure for setting up high-throughput nanolitre crystallization experiments. I. Protocol design and validation

• Published in: Journal of applied crystallography Vol. 36; no. 2; pp. 308 - 314
• Main Authors: Walter, T. S., Diprose, J., Brown, J., Pickford, M., Owens, R. J., Stuart, D. I., Harlos, K.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.04.2003; Blackwell
• Subjects: Biological and medical sciences; Cross-disciplinary physics: materials science; rheology; crystallization; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Growth from solutions; In solution. Condensed state. Thin layers; ink-jet technology; macromolecular crystallography; Materials science; Methods of crystal growth; physics of crystal growth; Molecular biophysics; Physico-chemical properties of biomolecules; Physics; protein crystallography; Methodology; Crystallization; Instrumentation; Biological macromolecule; Evaporation; Experimental protocol; Robotics; Proteins; Nanometer scale; Monocrystals; Sample preparation; Automatic system; Crystal growth from solutions; Droplets; Sitting drop method
• ISSN: 1600-5767; 0021-8898; 1600-5767
• DOI: 10.1107/S0021889803001997

A protocol for setting up nanolitre sitting‐drop vapour‐diffusion experiments is reported. The procedure uses standard crystallization screening kits and 96‐well crystallization plates. Reservoir solutions are transferred from 96‐deep‐well blocks to crystallization plates in a single step with a Robbins‐Hydra pipettor. Nanolitre droplets of protein as well as reservoir solution are dispensed by a Cartesian pipetting instrument. Experiments have been carried out to characterize the performance of this instrument. Adaptations to the Cartesian, which include an anti‐evaporation cover plate, are described and tested. The protocol was designed for a high‐throughput facility, but can be used in any standard crystallography laboratory.