pims at the oppf jon diprose embo course ebi, 23/09/2008

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.

■ 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/

Construct Design

Cloning

Purification

Crystal Growth

Crystallography…

OPTIC

Nautilus

PlateDB

Large-scaleExpression

Crystallogenesis

The OPPF PipelineBioinformatics

PCRSmall-scaleExpression

Characterization

Construct Design

Cloning

Purification

Crystal Growth

Crystallography…

OPTIC

xtalPiMS

Large-scaleExpression

Crystallogenesis

The OPPF PipelineBioinformatics

PCRSmall-scaleExpression

Characterization

Overview

■ The OPPF

■ Workflows and Protocols

■ Using PiMS at the OPPF

Workflows

Workflows in PiMS

PiMS Protocols

OPPF Protocols

Overview

■ The OPPF

■ Workflows and Protocols

■ Using PiMS at the OPPF

PCR plate design

■ Plates designed in a spreadsheet template

Primer order plate

■

Primer order form

■

Operon order form page 1

■

Operon order form page 2

Plate experiment workflow

Rescue and cleanup workflow

Scoring PCR plates

Scoring PCR plates

Scoring an expression plate

Scoring an expression plate

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

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

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