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Repeated Use of Qiagen Columns in Large-Scale Preparation of Plasmid DNA

  • Derek Gregory
  • Ricardo E. Tascon
  • Douglas B. Lowrie
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 29)

Abstract

The preparation of large amounts of high-purity intact plasmid DNA is a significant expense in DNA vaccine research. In our laboratory, mice are typically each immunized by injection of 100 μg on four occasions, so that an experiment with 50 mice requires 20 mg DNA as a minimum. The Qiagen tip-10,000 (Giga) column is intended to deliver up to 10 mg of high copy-number plasmid DNA from Escherichia coli. We have found that by repeatedly regenerating the column and reapplying the flowthrough volume of DNA extract, we can readily obtain 2–4-fold increased yields, up to 40 mg, from the standard 2.5 L broth culture.

Keywords

Escherichia Coli Agar Plate Sterile Distil Water Broth Culture Flask Shaker 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Reference

  1. 1.
    Tascon, R. E., Colston, M. J., Ragno, S., Stavropoulos, E., Gregory, D., and Lowrie, D. B. (1996) Vaccination against tuberculosis by DNA injection. Nat. Med. 2, 888–892.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Derek Gregory
    • 1
  • Ricardo E. Tascon
    • 1
  • Douglas B. Lowrie
    • 2
  1. 1.National Institute for Medical ResearchLondonUK
  2. 2.Mycobacterial Research LaboratoryLondonUK

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