Site-Directed Mutagenesis Using Double-Stranded Plasmid DNA Templates

  • Jeffrey Braman
  • Carol Papworth
  • Alan Greener
Part of the Springer Protocols Handbooks book series (SPH)


In vitro site-directed mutagenesis is an invaluable technique for studying protein structure-function relationships, gene expression and vector modification. Several methods for performing this technique have appeared in the literature (1, 2, 3, 4, 5). These procedures generally require multiple enzymatic steps, specialized vectors, and convenient restriction sites or subcloning of the sequence of DNA to be mutated into a bacteriophage vector like M13 to produce and recover single-stranded DNA. These manipulations present major limitations to the routine use of these methods because they are time consuming and tedious.


Competent Cell Control Reaction Experimental Reaction Polypropylene Tube Heat Pulse 
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Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Jeffrey Braman
    • 1
  • Carol Papworth
    • 1
  • Alan Greener
    • 1
  1. 1.StratageneLa Jolla

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