GeneArt™ Site-Directed Mutagenesis PLUS System

We do not offer the DNA methylase from GeneArt Site-Directed Mutagenesis kits as a separate product.

During the first cycle of PCR on the circular template, the polymerase will generate two linear products that are able to anneal and undergo primer extension in the next cycle. PCR products that incorporate the mutation at both ends will accumulate in subsequent cycles. After PCR, the recombination reaction results in circularization of the PCR product which enhances the mutagenesis efficiency.

The kits are almost the same, but the PLUS kit contains an improved 2X GeneArt enzyme mix with protocols optimized for multi-site mutagenesis (up to 3 sites in plasmids up to 14 kb). The PLUS kit can also be used for single-site mutations of up to 25 nucleotides, while the original kit can be used for single substitution/deletion/insertions up to 12 nucleotides in plasmids up to 14 kb. A web tool is available for primer design that can also be used for the original kit.

Yes. Add “single variant” as a new process, then choose an “upstream service” which is: gene synthesis, already at Geneart, or I will provide this master gene. If the customer is providing their own gene, they will need to fill in the field for the complete sequence of their vector + insert. If it is a plasmid only, they will put the mutagenized area as the “insert” and the rest of the plasmid as the “vector”. The “insert” can only be up to 4kb long.

Yes, 25X SAM is not stable and will lose activity within a few hours. We recommend creating a new dilution of SAM from the 200X SAM supplied in the kit each time you perform a mutagenesis procedure.