| Format | Phusion site directed mutagenisis |
| RNase inhibitor included | No |
| Hot-Start | Yes |
| High fidelity | Yes |
| Long distance (>5 kb) | Yes |
| Multiplex | No |
Learn more about this product
Thermo Scientific's Phusion Site-Directed Mutagenesis Kit provides reagents for 20 reactions to introduce point mutations, insertions or deletions into plasmid DNA. The method uses phosphorylated primers to amplify the complete plasmid while incorporating the required sequence change. The resulting linear PCR product is circularized in a five-minute ligation reaction with the included T4 DNA Ligase, after which the plasmid can be transformed into competent E. coli cells.
The workflow does not require special vectors, restriction sites or a particular methylation status, and the starting template does not need to be destroyed in a separate step. Phusion Hot Start II High-Fidelity DNA Polymerase helps minimize unwanted secondary mutations and supports amplification of plasmids up to 10 kb. Its hot-start modification limits non-specific amplification and unwanted primer degradation before the first PCR cycle. No purification is required before or after the ligation step, and the resulting plasmid is compatible with all strains of competent E. coli cells.