Site-directed, Ligase-Independent Mutagenesis (SLIM) for highly efficient mutagenesis of plasmids greater than 8kb

J Microbiol Methods. 2008 May;73(2):195-8. doi: 10.1016/j.mimet.2008.02.013. Epub 2008 Feb 26.


Modifying the Site-directed, Ligase-Independent Mutagenesis (SLIM) protocol from a single reaction mode to a two-reaction mode enables highly efficient mutagenesis of plasmid constructs that exceed 8kb. This modified approach reduces the complexity of the PCR step and is optimised for generation of heteroduplexes from long PCR products. The two-reaction mode SLIM has 92% efficiency.

MeSH terms

  • DNA, Bacterial / genetics*
  • Ligases / metabolism
  • Mutagenesis, Site-Directed / methods*
  • Plasmids*
  • Polymerase Chain Reaction / methods


  • DNA, Bacterial
  • Ligases