title: Site-directed, Ligase-Independent Mutagenesis (SLIM) for highly efficient mutagenesis of plasmids greater than 8kb.
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.
authors.affiliation: School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia. j.chiu@unsw.edu.au
authors.forename: Joyce
authors.initials: J
authors.lastname: Chiu
authors.forename: Daniel
authors.initials: D
authors.lastname: Tillett
authors.forename: Ian W
authors.initials: IW
authors.lastname: Dawes
authors.forename: Paul E
authors.initials: PE
authors.lastname: March
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics*; D008025:Ligases / Q000378:metabolism; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D004269:DNA, Bacterial; D008025:Ligases