Molecular Biotechnology

, Volume 29, Issue 3, pp 225–232 | Cite as

A single-stage polymerase-based protocol for the introduction of deletions and insertions without subcloning

  • John C. Salerno
  • Rachel J. Jones
  • Eda Erdogan
  • Susan M. E. Smith
Research Protocol


A single-stage polymerase-based procedure is described that allows extensive modifications of DNA. The version described here uses the QuikChange Site-Directed Mutagenesis System kit supplied by Stratagene. The original protocol is replaced by a single-stage method in which linear production of complementary strands is accomplished in separate single primer reactions. This has proved effective in introducting insertions and deletions into large gene/vector combinations without subcloning.

Index Entries

Linear amplification site-directed mutagenesis 


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Copyright information

© Humana Press Inc 2005

Authors and Affiliations

  • John C. Salerno
    • 1
  • Rachel J. Jones
    • 1
  • Eda Erdogan
    • 1
  • Susan M. E. Smith
    • 1
  1. 1.Biology DepartmentRensselaer Polytechnic Institute, Science CenterTroy

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