• Primers anneal to the DNA template, mutant strands are synthesized with 2×TransStart® FastPfu PCR SuperMix.
• In vitro digestion of non-mutated parental plasmid (methylated plasmid) with DMT enzyme and in vivo degradation of non-mutated parental plasmid (methylated plasmid) with DMT Chemically Competent Cell, so as to efficiently select mutant clones.
Highlight
• Mutation sites on both primers to improve mutation efficiency.
• Partially overlapping primers for exponential DNA amplification.
• Fast (4 kb/min) and high fidelity (54-fold fidelity as compared to EasyTaq® DNA Polymerase) 2xTransStart® FastPfu PCR SuperMix for DNA amplification.
• Double digestions (in vitro and in vivo) of parental plasmids to enhance mutation efficiency.
Storage
Competent Cell at -70°C for six months; others at -20°C for two years