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Protein Expression Factory

The Expression Factory (EF)- located in UCD Conway Institute is a microplate based automated system controlled by software that facilitates and monitors biochemical and molecular biology processes.

  • The maximum throughput is 96x4 samples at one time and the EF can facilitate primer design, PCR, subcloning into expression vectors, transformation, expression, lysis and purification of dozens of proteins in parallel. The system can use bacteria and cell-free systems for expression.
  • It can be used as an automated cloning system for multiple vectors with different affinity purification tags to be used in different expression systems such as mammalian cells, yeast, insect cells and others.
  • The protocols can also include restriction/ligation, Topo, as well as Gateway (Invitrogen), and Creator (Clontech) recombination systems to add flexibility to different cloning strategies.
  • It can generate dozens of mutants for a given protein/gene.
  • Compared to multiple subcloning and protein purifications at the bench, significant advantages of the EF include strategic support and training, software support, minimised risks and errors and throughput and speed

Services:

  • PCR (including primer design)
  • Gateway BP and LR reactions and additional cloning techniques
  • Growth of E.coli cells
  • Plasmid preparation (minipreps)
  • Restriction digestion (single and double enzyme digestion)
  • SAP and CIP treatment
  • Klenow treatment
  • Ligation
  • Transformation of plasmids into competent E.coli cells
  • Cell growth monitoring for E.coli by OD (600nm) and protein expression by fluorescence (GFP)
  • Expression culture induction
  • Cell lysis with detergent
  • Separation of soluble and insoluble E.coli cell lysis fractions by centrifugation
  • Sample preparation for SDS PAGE analysis
  • Solubilisation of insoluble inclusion bodies
  • Purification of proteins from fractions under native or denaturing conditions
  • On-column refolding and purification of denatured proteins on a nickel affinity matrix
  • Preparation of transposed baculovirus DNA for use in insect expression cultures (BAC to BAC system)
  • Native protein purification on pre-made or commercial resins.
  • Insoluble proteins may be solubilised and purified using automated affinity chromatography.

Instrumentation:

  • CRS robotic arm
  • Sias Xantas liquid handling system comprising of 8 channel pipettes
  • Automated plate handling
  • Four plate positon thermocyclers
  • Cytomat for cell growth and monitoring
  • Sias two plate centrifuge system
  • Filtration station
  • ‘Smart Trough’ system for purification and recovery

To register your interest, contact:

If you are looking for a quotation for using the Expression factory, please contact us by email and describe your project. We will send you a detailed estimate of the cost and the timeframes.

Check out our next associated technology: High Content Analysis

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