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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

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CBS

CRYSTALLISATION

The crystallization platform of the CBS offers access to robotized methods for crystallization of macromolecules (mainly proteins) using the vapour diffusion techniques. 

Sample characterization is a prerequisite before crystallization. Unless it has already been performed, samples will be analyzed by native and SDS-PAGE, as well as dynamic light scattering and thermal shift assay.

Crystallization is first attempted using general screens from commercial sources as well as specific screens made from our own stock solutions.

- Crystallization screens are prepared/dispensed with a Tecan RSP100 robot (to be updated),

- Sitting drops -from 100nl to 500nl- are performed by a Cartesian Honeybee 8+1 robot.

- Plates are stored and automatically imaged at 20°C in a Formulatrix Rock Imager system. Images are accessible to users and can be scored via a web-interface.

Hanging drop plates are then available for manual screen refinement and crystal growth optimization.

Plates can also be stored at 4° and 18°C and inspected through a Leica stereo-microscope.

The CBS is now equipped with G-Rob robot (Nat-X-ray) for in situ X-ray diffraction screening in order to monitor crystal quality and nature (salt/protein).

Devices:

A Tecan RSP100 robot  (to be updated) is used to prepare crystallization screens in 96-well deep-well blocks and to dispense the reservoirs.

A a Cartesian Honeybee 8+1 robot is used to set up the nanolitre droplets or macromolecule.