Fast Mutagenesis System & Fast MultiSite Mutagenesis System
Site-Directed Mutagenesis System is a simple and highly efficient method for in vitro site-directed mutagenesis. The entire procedure, including transformation, may be completed in less than 3 hours (when using a 3 kb plasmid). This unique system can generate base substitutions, deletions, or insertions of up to 12 nucleotides in DNA plasmids of up to 14 kb from any source, with no specialized vectors or restriction sites required. The high efficiency (>90% mutagenesis efficiency, depending on the size of the plasmid used) and simplified protocols of this kit allow for the generation of site-directed mutants. Two complementary mutagenic oligonucleotide primers with centrally located mutation sites are required to generate a mutation site, and the DNA methylation and amplification steps are combined in a single reaction. No in vitro digestion step is required after the mutagenesis reaction and no purification step is required after methylation or mutagenesis.
Applications
In vitro site-directed mutagenesis can be used to:
• Study protein function
• Identify enzyme active sites
• Design new proteins
The kits are almost the same, but the MULTISITE kit contains an improved 2X enzyme mix with protocols optimized for multi-site mutagenesis (up to 3 sites). A web tool was also introduced for primer design, but this can also be used for the original kit.
Workflow of Directed Mutagenesis System