Phusion^® Site-Directed Mutagenesis Kit

A fast and reliable system for generating mutations

Description

The Phusion® Site-Directed Mutagenesis Kit is a versatile and efficient tool for introducing point mutations, insertions or deletions in any type of plasmid DNA. 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 circularized in a 5-minute ligation reaction with Quick T4 DNA Ligase. The resulting plasmid can be then transformed into any competent E. coli cells.

General information about Phusion DNA Polymerases, their advantages and applications can be found on Phusion DNA Polymerase products page.

The Phusion^® Site-Directed Mutagenesis Protocol

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Advantages

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
Due to the high fidelity of Phusion® Hot Start DNA Polymerase unwanted secondary mutations are minimized
Amplification of large plasmids up to 10 kb
The hot start modification in the polymerase prevents the amplification of non-specific products and unwanted degradation of primers prior to first cycle of PCR
Quick T4 DNA Ligase included in the kit; no purification steps before or after the ligation
Compatible with all strains of competent E. coli cells

The Phusion® Site-Directed Mutagenesis Kit includes Phusion Hot Start DNA Polymerase, 5x Phusion HF Buffer, dNTP mix, Quick T4 DNA Ligase (NEB), 2x Quick Ligation Buffer (NEB) and a Control Plasmid with Control Primer Mix.