PCR Method for Generating Multiple Mutations at Adjacent Sites

  • Jiri Adamec
Part of the Methods in Molecular Biology™ book series (MIMB, volume 192)


Site-directed mutagenesis is a commonly used tool for identifying the role of specific amino acids in the structure and function of proteins. Various methods of in vitro mutagenesis have been described and are widely used for introducing modified coding sequences (1-7). In comparison, polymerase chain reaction (PCR)-based methods (1-6) are generally faster and more efficient than non-PCR-based methods (7). On the other hand, some PCR-based methods require two or more primers for each round of mutagenesis, whereas others need a single very long oligonucleotide or two round of PCR for introducing one mutation (5-7). These all can increase the cost of mutagenesis.


Polymerase Chain Reaction Polymerase Chain Reaction Product Polymerase Chain Reaction Fragment Mutagenic Primer QIAprep Spin 
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Copyright information

© Humana Press Inc. 2002

Authors and Affiliations

  • Jiri Adamec
    • 1
  1. 1.Mayo Clinic and FoundationRochester

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