期刊
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
卷 61, 期 -, 页码 651-657出版社
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S0907444905007808
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Crystallization trials at the Division of Structural Biology in Oxford are now almost exclusively carried out using a high- throughput workflow implemented in the Oxford Protein Production Facility. Initial crystallization screening is based on nanolitre- scale sitting- drop vapour- diffusion experiments ( typically 100 nl of protein plus 100 nl of reservoir solution per droplet) which use standard crystallization screening kits and 96- well crystallization plates. For 294 K crystallization trials the barcoded crystallization plates are entered into an automated storage system with a fully integrated imaging system. These plates are imaged in accordance with a pre- programmed schedule and the resulting digital data for each droplet are harvested into a laboratory information-management system ( LIMS), scored by crystal recognition software and displayed for user analysis via a web- based interface. Currently, storage for trials at 277 K is not automated and for imaging the crystallization plates are fed by hand into an imaging system from which the data enter the LIMS. The workflow includes two procedures for nanolitre- scale optimization of crystallization conditions: ( i) a protocol for variation of pH, reservoir dilution and protein: reservoir ratio and ( ii) an additive screen. Experience based on 592 crystallization projects is reported.
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