Thomas S. Walter1, Jonathan M. Diprose1, James Brown1, Mike G. Pickford1, Raymond J. Owens1, David I. Stuart1, Karl Harlos1
1Oxford Protein Production Facility and The Division of Structural Biology, The Henry Wellcome Building of Genomic Medicine, Roosevelt Drive, Headington, Oxford OX3 7BN, UK
Tóm tắt
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.
