Thermo Scientific™ Phusion™ Site-Directed Mutagenesis Kit with DH10B Competent Cells

Description

• 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 changes
  • The 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 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
  • Phusion Hot Start II High Fidelity DNA Polymerase minimizes unwanted secondary mutations\
  • Amplification of large plasmids up to 10 kb
  • Hot start modification of the polymerase prevents amplification of non-specific products and unwanted degradation of primers prior to first cycle of PCR
  • T4 DNA Ligase included in the kit; no purification steps before or after ligation
• DH10B Competent Cells
  • High efficiency, chemically competent E.coli cells
  • The DH10B strain is suitable for cloning DNA containing methylcytosine and methyladenine, allowing both prokaryotic and eukaryotic genomic DNA to be cloned efficiently
  • These cells are ideal for the construction of cDNA libraries using plasmid-derived vectors
  • Genotype: F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ (ara-leu)7697 galU galK λ– rpsL(StrR) nupG