GeneArt™ Site-Directed Mutagenesis System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Create a fresh dilution of 25X SAM from the kit-supplied 200X SAM in sterile, distilled water each time you perform the mutagenesis procedure - 25X SAM is not stable, and loses activity within a few hours after preparation

Protocol tips
- Create a fresh dilution of 25X SAM from the kit-supplied 200X SAM in sterile, distilled water each time you perform the mutagenesis procedure - 25X SAM is not stable, and loses activity within a few hours after preparation

Publication protocol

Plasmids
The transfer plasmids for the UL23 gene-kanamycin cassette were constructed by amplifying the native gene from the BAC HSV1(17+)Lox25,35 followed by cloning of the SalI and XmaJI digested amplicon into the plasmid pCeu248. The kanamycin resistance gene was amplified from pEPkan-S26 and ligated into the unique BglII-site resulting in the plasmid pCeu2-UL23(kanr). Based on this plasmid, nucleotide substitutions were introduced by the GeneArt site-directed mutagenesis system kit (Thermo Fisher Scientific, Waltham, MA, USA). The cloning strategy is depicted in the Supplementary Figure, and the oligonucleotides used for cloning and site-directed mutagenesis are provided in the Supplementary table.

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Reviews

Discussion

Papers

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for GeneArt™ Site-Directed Mutagenesis System below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Site Directed Mutagenesis (SDM) Monkey - Point mutation Vero UL23 thymidine kinase using GeneArt™ Site-Directed Mutagenesis System from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.