NKI Crystallisation Facility

The Crystallisation Facility at The Netherlands Cancer Institute (NKI) is part of the Structural Biology groups headed by Anastassis Perrakis and Titia Sixma. The facility provides a service for crystallisation of biomolecular samples including proteins, DNA, RNA and macromolecular complexes. Crystallisation service is offered only to people working at projects within the NKI or to collaborating research programs. The facility is also available for members of the European Consortia 3D-Repertoire and RUBICON.

3D-Repertoire is an integrated project funded by the European Commission under Framework 6 starting from 1st February 2005 and ending on 31st July 2009. 3D-Repertoire aims to resolve structures for all amenable protein complexes from budding yeast (or where necessary equivalents from other species).

RUBICON is a "Network of Excellence" funded by the Framework Programme 6 of the European Commission over a five years period (January 2006 - December 2010). RUBICON will provide a European forum for research on the molecular principles and regulatory roles of protein modifications by linkage of ubiquitin and ubiquitin-like molecules and will address the involvement of such modifications in major human diseases such as acute and chronic inflammation, neurodegeneration and cancer.

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  CRYSTALLISATION EQUIPMENT

Hydra II

Mosquito

The crystallisation facility at the NKI is equipped with state-of-the-art robotics including a Hydra II liquid handling robot for transferring crystallant solutions from a 96-Deep Well block to the reservoir of a crystallisation plate and a Mosquito nanodrop dispenser for preparation of nanoliter drops for vapour diffusion crystallisation experiments in 96-well format. Our custom setup prepares 200 nanoliter drops (100 nl sample + 100 nl crystallant solution) and requires only 28 microliter of sample for screening 96 crystallisation conditions. Crystallisation trials can be set up against a variety of commercially available screens, including PACT, JCSG+, Hampton, Wizard and Jena screens.

The facility also hosts a TOPAZ microfluidics system for free interface diffusion (FID) crystallisation experiments at nanoliter scale. This crystallisation technique is different from vapour diffusion and can be used as an additional screening method for identification of crystallisation conditions. This system requires only 1.5 nanoliter of sample for screening 96 conditions. There are 5 screens available that have been optimized for the microfluidics (Optimix 1 to 5 from Fluidigm), however also 'conventional' vapour diffusion screens can be used.

Reagent chamber volume: 2.55nL
Protein chamber volume: 0.75nL

Crystallisation trials are monitored by the Crystal Farm, an automated protein crystallisation storage and imaging system. Images of the crystallisation drops are automatically taken according to a user-defined schedule and can be viewed through a web-interface.


  CONTACT
Patrick Celie (PhD) 
Division of Molecular Carcinogenesis 
Plesmanlaan 121 
1066 CX  Amsterdam 
The Netherlands 
Telephone (office): + 31 20 512 2053
Telephone (lab): + 31 20 512 1964
Fax: + 31 20 512 1954