Thermo Fisher, F542, Phusion™ Site-Directed Mutagenesis Kit with DH10B Competent Cells

Quantity: 20 Reactions
Unit Size: Each
• This package includes a Phusion Site-Directed Mutagenesis Kit and DH10B Competent Cells that are optimized for use with the kit
• Phusion Site-Directed Mutagenesis Kit:With this kit the entire plasmid is amplified using phosphorylated primers that introduce the desired changesThe amplified, linear PCR product, containing the desired mutation, is circularized in a 5-minute ligation reaction with T4 DNA Ligase. The resulting plasmid can be then transformed into the E.coli cells. DH10B Competent Cells are recommended for use with the kit.The optimal annealing temperature for Phusion DNA polymerases may differ significantly from that of Taq-based polymerases. For optimal results start by accurately calculating your Tm with our Tm calculator.Robust and reliable exponential amplification methodNo requirements, such as special vectors, restriction sites, or methylation status for the target plasmidNo need to destroy the starting template in a separate stepPhusion Hot Start II High Fidelity DNA Polymerase minimizes unwanted secondary mutations\Amplification of large plasmids up to 10 kbHot start modification of the polymerase prevents amplification of non-specific products and unwanted degradation of primers prior to first cycle of PCRT4 DNA Ligase included in the kit; no purification steps before or after ligation
• DH10B Competent CellsHigh efficiency, chemically competent E.coli cellsThe DH10B strain is suitable for cloning DNA containing methylcytosine and methyladenine, allowing both prokaryotic and eukaryotic genomic DNA to be cloned efficientlyThese cells are ideal for the construction of cDNA libraries using plasmid-derived vectorsGenotype: F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ (ara-leu)7697 galU galK λ– rpsL(StrR) nupG