LiClone™ Fast Mutagenesis Kit (10 rxns)

LiClone™ Fast Mutagenesis Kit is a site-directed mutagenesis system based on the LiClone homologous recombination technology for introducing single or double point mutations in plasmids. The kit integrates the LiTaq Super-Fidelity DNA Polymerase amplification system and the LiClone rapid cloning system. With superior fidelity, LiTaq 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 LiClone rapid cloning system replaces the conventional annealing-cyclization reactions with the more efficient homologous recombination reaction.

Site-directed mutagenesis using the LiClone™ Fast Mutagenesis Kit 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 DpnI-digested product can be used directly in the recombination reaction without DNA purification.
 

Key Features

• Low template usage: Robust and reliable exponential amplification method.

• High-fidelity enzyme: LiTaq Super-Fidelity DNA Polymerase (~128× Taq) minimizes unwanted secondary mutations.

• Simple operation: Amplified product can be directly used in the recombination reaction after being digested with DpnI.

• Widely applicable: Suitable for amplification of any site in the plasmid within 20 kb. 
 

Applications

1. Mutation: 1 - 2 mutation sites (multiple bases within 50 bp apart can be counted as 1 mutation site)

2. Micro-fragment insertion: insertion of a specific sequence of no more than 50 bp at any site of the vector

3. Deletion: deletion of a specific sequence on a vector
 

Components

1. 2× LiClone buffer: 1.25 ml
2. dNTP mix (10 mM each): 20 µll
3. LiTaq Super-Fidelity DNA Polymerase: 20 µl
4. DpnI (10 U/µl): 20 µl
5. 5× CE II buffer: 40 µll
6. Exnase: 20 µll
 

Storage

Store at at -20°C.