The method works first by having primers anneal to the DNA template. Mutant strands are then synthesized with 2×TransStart® FastPfu PCR SuperMix with high fidelity. In the next step, non-mutated parental plasmid (methylated plasmid) is digested in vitro with DMT enzyme and in vivo further digested within DMT Chemically Competent Cell.
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• 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.
Competent Cell at -70°C for six months; others at -20°C for two years
FM111-01 (10 rxns)
FM111-02 (20 rxns)
2×TransStart® FastPfu PCR SuperMix
DMT Enzyme (10 units/μl)
DMT Chemically Competent Cell
SControl Plasmid (5 ng/µl)
SControl Primers (10 µM )
Papers citing this product:
|Whole-genome sequencing of the snub-nosed monkey provides insights into folivory and evolutionary history.||Nature genetics||29.648||Xuming Zhou, et al.||2014 Dec||The original link|
|Crystal structure of PXY-TDIF complex reveals a conserved recognition mechanism among CLE peptide-receptor pairs||Cell Research||14.812||Heqiao Zhang, et al. Ministry of Education Key Lab||2016 jan||The original link|
|Maternal xNorrin, a canonical Wnt signaling agonist and TGF-β antagonist, controls early neuroectoderm specification in Xenopus||PloS Biology||12.69||Xu S, et al. State Key Laboratory of Molecular Dev||2012||The original link|
|PGE2 Promotes Hepatic Biliary Cholesterol Excretion by EP3-mediated HNF4α/CYP7A1 Pathway in Mice||hepatology||11.711||Shuai Yan,et al.||2016Dec||The original link|
|Plastid-nucleus communication involves calcium-modulated MAPK signalling||Nature communications||11.329||Hailong Guo, et al. Photosynthesis Research Center||2016 jun||The original link|