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PiMS at the OPPF

PiMS at the OPPF. Jon Diprose EMBO Course EBI, 23/09/2008. Overview. The OPPF Workflows and Protocols Using PiMS at the OPPF. Aims of the OPPF. To develop technology for high throughput protein production and crystallisation.

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PiMS at the OPPF

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  1. PiMS at the OPPF Jon Diprose EMBO Course EBI, 23/09/2008

  2. Overview • The OPPF • Workflows and Protocols • Using PiMS at the OPPF

  3. Aims of the OPPF • To develop technology for high throughput protein production and crystallisation. • To apply this technology to challenging biomedical problems through the development of a portfolio of collaborative projects. • To make the technology available to the wider structural biology community. http://www.oppf.ox.ac.uk/

  4. Crystallogenesis Crystallography… The OPPF Pipeline Bioinformatics Construct Design Small-scale Expression PCR Cloning Large-scale Expression Characterization Purification OPTIC Crystal Growth Nautilus PlateDB

  5. Crystallogenesis Crystallography… The OPPF Pipeline Bioinformatics Construct Design Small-scale Expression PCR Cloning Large-scale Expression Characterization Purification OPTIC Crystal Growth xtalPiMS

  6. Overview • The OPPF • Workflows and Protocols • Using PiMS at the OPPF

  7. Workflows

  8. Workflows in PiMS

  9. PiMS Protocols

  10. OPPF Protocols

  11. Overview • The OPPF • Workflows and Protocols • Using PiMS at the OPPF

  12. PCR plate design • Plates designed in a spreadsheet template

  13. Primer order plate

  14. Primer order form

  15. Operon order form page 1

  16. Operon order form page 2

  17. Plate experiment workflow

  18. Rescue and cleanup workflow

  19. Scoring PCR plates

  20. Scoring PCR plates

  21. Scoring an expression plate

  22. Scoring an expression plate

  23. Purification protocols • 9 protocols to match Akta programmes • Single column protocols • Affinity with gradient elution • Batch affinity • Desalt • Gel filtration • Ion exchange • Two column protocols • Affinity desalt • Affinity gel filtration • Mammalian affinity gel filtration • Mammalian affinity desalt

  24. PiMS usage • PiMS has been in use at the OPPF for 18 months • Used exclusively to record wet-lab work this year • 221 Targets • 134 Plates • 544 Simple experiments

  25. Acknowledgements • Dave Stuart (PI) • Ray Owens (Project Manager) • Robert Esnouf • Jon Diprose • Marc Savitsky (PiMS) • Ian Berry (now Evotec) • Joanne Nettleship • Louise Bird • Kim Henrick (PI) • Chris Morris (Project Manager) • Ed Daniels • Susy Griffiths • Bill Lin • Jo van Niekirk (now Ingres) • Anne Pajon • Peter Troshin

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