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Nucleic Acids pp 191-195 | Cite as

Small-Scale Plasmid DNA Preparation

  • J. W. Dale
  • P. J. Greenaway
Part of the Methods in Molecular Biology book series (MIMB, volume 2)

Abstract

For the initial characterization of a recombinant plasmid, it is necessary to determine the size of the plasmid or, preferably, the size and characteristics of the insert itself. A method is therefore required for the simultaneous preparation, from a number of isolates, of plasmid DNA in a state sufficiently pure for restriction enzyme digestion. The requirements of such a procedure are:
  1. (i)

    A simple method for rapid lysis of the bacterial cells.

     
  2. (ii)

    Separation of plasmid from chromosomal DNA.

     
  3. (iii)

    Removal of proteins and of other components of the cells that might interfere with restriction enzyme treatment.

     
  4. (iv)

    Removal of detergents, salts, etc. used in the process.

     

Keywords

Sodium Dodecyl Sulfate Ethanol Precipitation Microfuge Tube Rapid Lysis Restriction Enzyme Treatment 
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.

References

  1. 1.
    Birnboim, H. C, and Doly, J. (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl. Acids Res. 7, 1513–1523.PubMedCrossRefGoogle Scholar
  2. 2.
    Wheatcroft, R., and Williams, P. A. (1981) Rapid methods for the study of both stable and unstable plasmids in Pseudomonas. J. Gen. Microbiol. 124, 433–437.PubMedGoogle Scholar

Copyright information

© The Humana Press Inc. 1984

Authors and Affiliations

  • J. W. Dale
    • 1
    • 2
  • P. J. Greenaway
    • 1
    • 2
  1. 1.Department of MicrobiologyUniversity of SurreyGuildford
  2. 2.Molecular Genetics LaboratoryPHLS Centre for Applied Microbiology and Research, Porton DownSalisburyUnited Kindgom

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