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Microseeding Matrix Screening and other developments in automatic protein crystallization

Microseeding Matrix Screening and other developments in automatic protein crystallization Patrick Shaw Stewart Douglas Instruments Limited (near Oxford, UK): Peter Baldock, Patrick Shaw Stewart, James Smith. Microseeding in screening experiments. Allan D’Arcy Novartis, Basle

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Microseeding Matrix Screening and other developments in automatic protein crystallization

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  1. Microseeding Matrix Screening and other developments in automatic protein crystallization Patrick Shaw Stewart Douglas Instruments Limited (near Oxford, UK): Peter Baldock, Patrick Shaw Stewart, James Smith

  2. Microseeding in screening experiments Allan D’Arcy Novartis, Basle 2006 ‘Matrix-seeding script’ An automated microseed matrix-screening method for protein crystallization. ActaCryst. (2007). D63, 550–554

  3. Phase diagram of a protein precipitate nucleation [Protein] metastable zone clear [Precipitant]

  4. Microseeding in screening experiments Gregory Ireton and Barry Stoddard. 'Microseed matrix screening to improve crystals of yeast cytosine deaminase'. Acta Crystallographica section D60 (2004), 601–605. With Ca2+ conditions they couldn’t get crystals without microseeding.

  5. Phase diagram of a protein precipitate [Protein] metastable zone clear [Precipitant]

  6. Microseeding in screening experiments • May explain why there are SO many hits with MMS • Can be used to CONTROL number of conditions

  7. Microseeding in screening experiments James Smith 2006 ‘Matrix-seeding script’

  8. Microseeding in screening experiments James Smith 2006 ‘Matrix-seeding script’

  9. Microseedinginscreeningexperiments • Lesley Haire, NIMR: • 6 hits in a screen, all poorly formed. • Seeding - 30 hits, including several well-formed crystals. • Jens-Christian Navaro Poulsen, University of Copenhagen • 1 hit in 288 wells • Seeding - 10 hits in 96 wells • Using one of those crystals for a second-generation seeding experiment - another 10 hits in 96 wells

  10. Microseedinginscreeningexperiments Noseeds Laura Cendron University of Padova

  11. Microseedinginscreeningexperiments Noseeds Seeds Laura Cendron University of Padova

  12. Phase diagram of a protein [Protein] [Seeds]

  13. Mini-optimization

  14. Under £24,000 Under $46,000

  15. Bullet points: • 100 + 100 nl sitting drop • Only 9.8 μl of protein to set up a 96-well plate • Additive screens • Microseeding • Mini-optimization and optimization

  16. Jonathan Hadden • 96-well plate takes 8 minutes • There is an evaporation shield

  17. Under £24,000 Under $46,000

  18. ORYX (Arabian) The Biblical Zoo in Jerusalem

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