Fast Mutagenesis Kits are site-directed mutagenesis system based on LiCloneTM rapid cloning technology. LiSparkTM Max Super-fidelity DNA Polymerase is included for amplification. This kit uses proprietary assembly mix and homologous recombination to seamlessly assemble up to five mutagenesis fragments.
• LiSparkTM Super-Fidelity DNA Polymerase provides high-fidelity PCR with the lowest mutation rate.
• LiSparkTM Super-Fidelity DNA Polymerase with excellent long fragment amplification capability can be widely used for any plasmid amplification within 20 kb.
• Amplification is carried out exponentially and the template usage is extremely low, which is beneficial to the complete degradation of the original methylation template.
• DpnI eliminates contamination of the original template.
• Rapid cloning system can efficiently cyclize PCR products.
• The amplified products can be directly used for recombinant reaction after digestion with DpnI.
• LiCloneTM Fast Mutagenesis Kit: 1-2 discontinuous sites (more than 50 bp apart) at one time on the target plasmid.
• LiCloneTM Fast MultiSite Mutagenesis Kit: 3-5 discontinuous sites (more than 50 bp apart) at one time on the target plasmid.
Choose the mutation sites A and B as the boundary to divide the plasmid into fragment AB and fragment BA. Design partially reverse complement primers contain at the two mutation sites.Amplify fragment AB (A Forward Primer and B Reverse Primer) and fragment BA (A Reverse Primer and B Forward Primer) with the original plasmid as template. (Step I). Amplification product was digested by DpnI (Step I), and then used in the recombination reaction (Step II). Recombination product was transformed directly to complete the double base site-directed mutagenesis (Step III).
|Product Name||Cat. #||Size||Price||Order|
|LiCloneTM Fast Mutagenesis Kit||M0011||20 rxns||$189|
|LiCloneTM Fast MultiSite Mutagenesis Kit||M0012||25 rxns||$399|