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Homemade Site Directed Mutagenesis of Whole Plasmids
Published on: May 11, 2009
Site-directed, Ligase-Independent Mutagenesis (SLIM) for highly efficient mutagenesis of plasmids greater than 8kb
Joyce Chiu1, Daniel Tillett, Ian W Dawes
1School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia. j.chiu@unsw.edu.au
Journal of Microbiological Methods
|April 5, 2008
Abstract:
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.

