GeneArt® Site-Saturation Mutagenesis | Thermo Fisher Scientific

Systematic Mutagenesis

The GeneArt® Site-Saturation Mutagenesis Service systematically replaces wild type amino acid coding sequences with sequences encoding all 19 non–wild type amino acids at the position(s) you specify. Also known as sequential permutation, site-saturation mutagenesis in combination with a powerful screening assay is the most systematic mutation strategy to identify amino acid substitutions that fulfill your protein engineering goals.

Once you’ve identified variants with beneficial substitutions, the GeneArt® Combinatorial Library Service can be used to create material for more targeted directed-evolution experiments to further optimize your protein.

Get Your Project Started

Please download the questionnaire to submit project information. For secure data transfer please register at our customer portal.

For further information regarding this service or the order process, please contact jpgene@lifetech.com.

Options for Site-Saturation Mutagenesis

We offer several options for sequential permutation to fit a wide array of research needs. You can choose to create variants at one or more codons, and receive constructs individually or pooled. When evaluating most of the possible 19 non–wild type amino acids is sufficient, we offer the GeneArt® Site-Saturation Mutagenesis Average 16 and GeneArt® Site-Saturation Mutagenesis Minimum 16 Services that provide individual clones encoding an average of 16 or a minimum of 16 substitute amino acids at each position, respectively.

Constructs supplied individually are 100% sequence-verified. Pooled constructs are bulk-sequenced to verify that the nucleotide content at the requested positions is degenerate and the nonmutated regions are intact
Variant constructs are subcloned into the vector of your choice

GeneArt® Site-Saturation Mutagenesis Pool of One Position	A mixture of all 19 non–wild type variants at one codon position Supplied as a pooled glycerol stock
GeneArt® Site-Saturation Mutagenesis Pool of All Positions	A mixture of all 19 non–wild type variants at more than one codon position Supplied as a pooled glycerol stock
GeneArt® Site-Saturation Mutagenesis Average 16	An average of 16 different non–wild type amino acids per position Supplied as individual glycerol stocks
GeneArt® Site-Saturation Mutagenesis Minimum 16	At least 16 different non–wild type amino acids per position Supplied as individual glycerol stocks
GeneArt® Site-Saturation Mutagenesis 19	All 19 different non–wild type amino acids per position Supplied as individual glycerol stocks

Applications

Identify beneficial or detrimental amino acid substitutions
Increase affinity, specificity, activity, heat stability, detergent tolerance, etc.
Change substrate specificity or enantioselectivity
Reduce homology between two related proteins (avoid IP issues)
Identify active sites or receptor-binding sites

Advantages

Systematic identification of beneficial amino acid substitutions—check every possible variant at each position
Structural information about the protein is not needed, but it can be beneficial
Fewer variants to screen—no oversampling is required when all clones are delivered in separate tubes
Fast delivery times—get your project moving quickly
Extremely cost-efficient

References

Bridge helix and trigger loop perturbations generate superactive RNA polymerases

For Research Use Only. Not for use in diagnostic procedures.

Stylesheet for Classic Wide Template adjustments