In vitro site-directed mutagenesis is an invaluable technique for characterizing the dynamic, complex relationships between protein structure and function, for studying gene expression elements, and for carrying out vector modification. Several approaches to this technique have been published, but these methods generally require single-stranded DNA (ssDNA) as the template and are labor intensive or technically difficult. Our QuikChange Lightning Site-Directed Mutagenesis Kit allows site specific mutation in virtually any double-stranded plasmid, thus eliminating the need for subcloning and for ssDNA rescue. In addition, the QuikChange Lightning Site-Directed Mutagenesis Kits do not require specialized vectors, unique restriction sites, multiple transformations or in vitro methylation treatment steps. The simple, rapid three-step generates mutants with high efficiency.
Using the most advanced high fidelity enzyme technology, the protocols have been accelerated while maintaining the highest accuracy for site-directed mutagenesis (see Figure 1). Exclusive to the QuikChange Lightning Site-Directed Mutagenesis Kit is a proprietary Pfu-based polymerase blend and the newly optimized Dpn I enzyme, which together allow for mutagenesis in approximately one hour, plus an overnight transformation.
Figure 1. Overview of the QuikChange Lightning Site-directed Mutagenesis Method.
The QuikChange Lightning Site-Directed Mutagenesis Kit delivers mutant plasmids up to three times faster than our original QuikChange kits, without losses in mutagenesis efficiency or accuracy. The kit has been optimized for mutagenesis of plasmids of up to 14 kb, allowing rapid, efficient, and accurate mutagenesis of small and large plasmids with a single kit.
QuikChange Lightning Multi Site-Directed Mutagenesis Kit
The QuikChange Lightning Multi Site-Directed Mutagenesis Kit offers the most rapid and reliable method for site-directed mutagenesis of plasmid DNA at up to five different sites simultaneously. Using the most advanced high fidelity enzyme technology, the protocols have been accelerated while maintaining the highest accuracy for site-directed mutagenesis. No specialized vectors or unique restriction sites are needed–virtually any plasmid of up to 8 kb is a suitable template. The rapid three-step procedure used for the QuikChange Lightning Multi kit introduces mutations at three different sites simultaneously in the 4-kb control plasmid with greater than 55% efficiency.
As with our original QuikChange Multi kit, a single mutagenic oligonucleotide is required to mutagenize each site, using a double-stranded DNA template. The kit’s exclusive enzyme blend features a unique Pfu Fusion-based DNA polymerase, providing high fidelity DNA synthesis. The rapid PCR cycling parameters of the unique enzyme blend, together with our newly optimized Dpn I enzyme, allow for multi-site mutagenesis in approximately two hours, plus an overnight transformation. The QuikChange Lightning Multi Site-Directed Mutagenesis Kit delivers results up to three times faster than our original multi-site mutagenesis kit (see Figure 2).
Figure 2. QuikChange Lightning Multi Saves Hours of Valuable Research Time.
For Research Use Only. Not for use in diagnostic procedures