AffiGEN® Express II Fast Mutagenesis Kit V2
AffiGEN® Express II Fast Mutagenesis Kit V2 is a site-directed mutagenesis system based on the ClonExpress homologous recombination technology for introducing single or double point mutations in plasmids. The kit integrates the Phanta Max Super-Fidelity DNA Polymerase amplification system and the ClonExpress rapid cloning system. With superior fidelity, Phanta Max significantly reduces the possibility of introducing novel mutations during amplification. Its excellent long fragment amplification ability is widely applicable to the amplification of any plasmid less than 20 kb. The ClonExpress rapid cloning system replaces the conventional annealing-cyclization reactions with the more efficient homologous recombination reaction. Site-directed mutagenesis using the Mut Express II Fast Mutagenesis Kit V2 allows a more flexible primer design and extremely low template input, thereby facilitating complete degradation of the initial methylated template. The highly optimized recombination reaction buffer and enhanced recombinase Exnase II significantly improve the efficiency of recombinant cyclization and tolerance to impurities. If the amplification product is highly specific, its Dpn I-digested product can be used directly in the recombination reaction without DNA purification.
Application:
- Mutation: 1 - 2 mutation sites (multiple bases within 50 bp apart can be counted as 1 mutation site)
- Micro-fragment insertion: insertion of a specific sequence of no more than 50 bp at any site of the vector
- Deletion: deletion of a specific sequence on a vector
| Components | 10 Reactions | 25 Reactions |
| 2 × Max Buffer | 1.25 mL | 1.25 mL |
| dNTP Mix (10 mM each) | 20 μL | 50 μL |
| dNTP Mix (10 mM each) Phanta Max Super-Fidelity DNA Polymerase | 20 μL | 50 μL |
| Dpn I (10 U/μl) | 20 μL | 50 μL |
| 5 × CE II Buffer | 40 μL | 100 μL |
| Buffer Exnase II | 20 μL | 50 μL |