Journal of Applied Crystallography

Volume 36, Part 2 (April 2003)

research papers

J. Appl. Cryst. (2003). 36, 308-314 [ doi:10.1107/S0021889803001997 ]

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

T. S. Walter, J. Diprose, J. Brown, M. Pickford, R. J. Owens, D. I. Stuart and K. Harlos

Abstract: 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.

Keywords: crystallization; macromolecular crystallography; protein crystallography; ink-jet technology.