Phusion® Site-Directed Mutagenesis Kit is a versatile and efficient tool for introducing point mutations, insertions, or deletions in any type of plasmid DNA.
- Robust and reliable exponential amplification method
- No requirements, such as special vectors, restriction sites, or methylation status for the target plasmid
- No need to destroy the starting template in a separate step
- Minimises unwanted secondary mutations
- Amplification of large plasmids up to 10 kb
- T4 DNA ligase included in the kit; no purification steps before or after ligation
- Compatible with all strains of competent E. coli cells
With this kit, the entire plasmid is amplified using phosphorylated primers that introduce the desired changes. The amplified, linear PCR product, containing the desired mutation, is circularised in a 5-minute ligation reaction with T4 DNA ligase. The resulting plasmid can be then transformed into any competent E. coli cells.
The optimal annealing temperature for Phusion DNA polymerases may differ significantly from that of Taq-based polymerases. Hot start modification of the polymerase prevents amplification of non-specific products and unwanted degradation of primers prior to first cycle of PCR.
Upozornení: For research use only. Not for use in diagnostic procedures.
