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Related Experiment Videos

High throughput methods for gene cloning and expression.

Lynda Dieckman1, Minyi Gu, Lucy Stols

  • 1Biosciences Division, Argonne National Laboratory, Argonne, IL 60439, USA.

Protein Expression and Purification
|June 20, 2002
PubMed
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This study presents an automated, high-throughput cloning process for producing bacterial expression clones. The method uses ligation-independent cloning, yielding 400 clones in 3 days for protein production and structural genomics.

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Structural Genomics

Background:

  • High-throughput production of bacterial expression clones is crucial for structural genomics and other applications.
  • Existing cloning methods can be time-consuming and labor-intensive.
  • Automated liquid handling systems offer potential for increased efficiency and throughput.

Purpose of the Study:

  • To develop and outline a high-throughput process for producing bacterial expression clones using automated liquid handlers.
  • To adapt the ligation-independent cloning (LIC) approach for automation.
  • To enable the simultaneous production of plasmids for diverse expression systems.

Main Methods:

  • Utilized automated liquid handlers for a series of interlinked liquid manipulation and incubation methods.

Related Experiment Videos

  • Employed the ligation-independent cloning (LIC) strategy for plasmid construction.
  • Integrated various stations of the automation system into a cohesive workflow.

    • Main Results:

    • Developed a 3-day protocol for bacterial expression clone production.
    • Achieved a linear throughput of 400 targets per production run.
    • Demonstrated the capability for simultaneous plasmid production for different expression systems.

    Conclusions:

    • The automated high-throughput cloning process significantly enhances the production of bacterial expression clones.
    • This method facilitates the generation of large numbers of clones for purified protein production.
    • The molecular protocols are applicable to various high-throughput strategies, including site-specific mutagenesis and protein interaction studies.