Accedi

Harwell Science and Innovation Campus

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

A Sample Preparation Pipeline for Microcrystals at the VMXm Beamline

Adam D. Crawshaw¹, Emma V. Beale^1,2, Anna J. Warren¹, Andrew Stallwood^1,3, Graham Duller¹, Jose Trincao¹, Gwyndaf Evans^1,4

¹Diamond Light Source Ltd, Harwell Science and Innovation Campus, ²Paul Scherrer Institut, ³Central Laser Facility, Science and Technologies Facilities Council, Harwell Science and Innovation Campus, ⁴Rosalind Franklin Institute, Harwell Science and Innovation Campus

The signal-to-noise ratio of data is one of the most important considerations in performing X-ray diffraction measurements from microcrystals. The VMXm beamline provides a low-noise environment and microbeam for such experiments. Here, we describe sample preparation methods for mounting and cooling microcrystals for VMXm and other microfocus macromolecular crystallography beamlines.

Biochemistry

Crystallization and In Situ Room Temperature Data Collection Using the Crystallization Facility at Harwell and Beamline VMXi, Diamond Light Source

James Sandy¹, Halina Mikolajek^1,2, Amy J. Thompson¹, Juan Sanchez-Weatherby¹, Michael A Hough^1,2

¹Diamond Light Source Ltd, Harwell Science and Innovation Campus, ²Research Complex at Harwell, Harwell Science and Innovation Campus

We present a protocol for the crystallization of proteins using the crystallization facility in the Research Complex at Harwell and subsequent in situ X-ray crystallographic data collection from crystals within the plates at Diamond's Versatile Macromolecular Crystallography in situ (VMXi) beamline. We describe sample requirements, crystallization protocols, and data collection guidelines.