Large scale purification of linear plasmid DNA for efficient high throughput cloning

Authors

  • Dr. Marjolaine Noirclerc-Savoye,

    Corresponding author
    1. CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    2. CNRS, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    3. Université Joseph Fourier, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    • Laboratoire d'Ingénierie des Macromolécules, Institut de Biologie Structurale, 41 rue Jules Horowitz, 38027 Grenoble, France, Fax: +33-4-3878-5494
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  • Benoit Gallet,

    1. CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    2. CNRS, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    3. Université Joseph Fourier, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
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  • Florent Bernaudat,

    1. CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    2. CNRS, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    3. Université Joseph Fourier, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
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  • Thierry Vernet

    1. CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    2. CNRS, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
    3. Université Joseph Fourier, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France
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Abstract

In this report we describe a rapid, simple, and efficient method for large-scale purification of linear plasmid DNA to answer demand from high-throughput gene cloning. The process is based on the separation of the linear vector from small DNA fragments by anion exchange chromatography. Gene cloning experiments by restriction/ligation or the In-Fusion(tm) technique confirmed the high quality of the linearized vector as 100% of the genes were successfully cloned.

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