Mutagenesis

Robust, high-efficiency systems and kits to streamline your workflow

  • Speed—time-to-results is typically less than 3 hours for 3 kb plasmid
  • Precision—alter up to 25 nucleotides (nt) when only one site is mutated
  • Flexibility—generate substitutions, deletions, or insertions in virtually any plasmid
  • Simplicityfree online tool  guides you through construct and primer design and generates final construct sequence for easy implementation

Which Site-Directed Mutagenesis System is Right for You?

 
1 site/plasmidyesyes
3 sites/plasmidyes
Mutagenesis Efficiency>90% for 1 site>90% for 1, 2, or 3 sites
# of MutationsUp to 25 nt mutations in 1 site/plasmid
  • Up to 25 nt mutations, up to 12 degenerate (single site)
  • Up to 3 nt mutations, up to 3 degenerate (multi site)
Time-to-ResultsTypically less than 3 hours for 3 kb plasmidTypically ~3 hours for 10 kb plasmid or smaller
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GeneArt® Site-Directed Mutagenesis Workflow

  Figure 1.  An overview of the workflow for GeneArt Site-directed Mutagenesis. The system relies on the inherent properties of DNA methylase, high fidelity DNA polymerase, recombination enzymes, and McrBC endonuclease.

High Mutagenesis Efficiency

 Multiple base mutagenesis is common, and we tested a 12 base substitution, insertion and deletion using a pUC19 plasmid. A random 12 base substitution was carried out within a single mutated primer. Alternatively, a random 12 base oligonucleotide containing a stop codon was inserted into wild type pUC19 plasmid, followed by deletion of the exact 12 bases to restore the wild type plasmid.

 

 The mutagenesis efficiency for a 12 base substitution, insertion or deletion was above 90%. The performance of GeneArt® Site-Directed Mutagenesis kit was comparable to the latest generation of kits from competitor 'Q'.

Take control of your mutagenesis experiments today!

Multisite-Directed Mutagenesis Efficiency

 Figure indicates the multisite-directed mutagenesis efficiency of 3 sites of 1 bp or 3 bp each in 5, 10, and 14 kb plasmids. In all cases the mutated sites (1 or 3 bp each) included one insertion, one deletion and one substitution. The performance of GeneArt® Site-Directed Mutagenesis PLUS System was comparable to the latest generation of multisite-directed mutagenesis kits from the competitor.

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