High-throughput in situ experimental phasing

DOI: 10.1107/S2059798320009109

Authors: Joshua M. Lawrence (Diamond Light Source) , Julien Orlans (Diamond Light Source; University of Lyon) , Gwyndaf Evans (Diamond Light Source) , Allen M. Orville (Diamond Light Source; Research Complex at Harwell) , James Foadi (Diamond Light Source) , Pierre Aller (Diamond Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acta Crystallographica Section D Structural Biology , VOL 76 , PAGES 790 - 801

State: Published (Approved)
Published: August 2020
Diamond Proposal Number(s): 14449 , 16818

Abstract: In this article, a new approach to experimental phasing for macromolecular crystallography (MX) at synchrotrons is introduced and described for the first time. It makes use of automated robotics applied to a multi-crystal framework in which human intervention is reduced to a minimum. Hundreds of samples are automatically soaked in heavy-atom solutions, using a Labcyte Inc. Echo 550 Liquid Handler, in a highly controlled and optimized fashion in order to generate derivatized and isomorphous crystals. Partial data sets obtained on MX beamlines using an in situ setup for data collection are processed with the aim of producing good-quality anomalous signal leading to successful experimental phasing.

Journal Keywords: in situ crystallography; experimental phasing; crystal soaking; high throughput; isomorphism

Subject Areas: Technique Development, Biology and Bio-materials


Instruments: I04-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography

Documents:
qh5065.pdf

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