A procedure for setting up high-throughput nanolitre crystallization experiments. II. Crystallization results

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Brown, J ; Walter, T S ; Carter, L ; Abrescia, N G A ; Aricescu, A R ; Batuwangala, T D ; Bird, L E ; Brown, N ; Chamberlain, P P ; Davis, S J ; Dubinina, E ; Endicott, J ; Fennelly, J A ; Gilbert, R J C ; Harkiolaki, M ; Hon, W-C ; Kimberley, F ; Love, C A ; Mancini, E J ; Manso-Sancho, R ; Nichols, C E ; Robinson, R A ; Sutton, G C ; Schueller, N ; Sleeman, M C ; Stewart-Jones, G B ; Vuong, M ; Welburn, J ; Zhang, Z ; Stammers, D K ... view all 34 authors (2003)

An initial tranche of results from day-to-day use of a robotic system for setting up 100 nl-scale vapour-diffusion sitting-drop protein crystallizations has been surveyed. The database of over 50 unrelated samples represents a snapshot of projects currently at the stage of crystallization trials in Oxford research groups and as such encompasses a broad range of proteins. The results indicate that the nanolitre-scale methodology consistently identifies more crystallization conditions than traditional hand-pipetting-style methods; however, in a number of cases successful scale-up is then problematic. Crystals grown in the initial 100 nl-scale drops have in the majority of cases allowed useful characterization of x-ray diffraction, either in-house or at synchrotron beamlines. For a significant number of projects, full x-ray diffraction data sets have been collected to 3 Å resolution or better (either in-house or at the synchrotron) from crystals grown at the 100 nl scale. To date, five structures have been determined by molecular replacement directly from such data and a further three from scale-up of conditions established at the nanolitre scale.
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