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