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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Papers
Check out relevant papers found by Labettor's AI that are relevant for performing Site Directed Mutagenesis (SDM) Monkey - Point mutation Vero UL23 thymidine kinase using GeneArt™ Site-Directed Mutagenesis System from Thermo Fisher Scientific.
Paper title
Recombinant herpes simplex virus type 1 strains with targeted mutations relevant for aciclovir susceptibility
Manufacturer protocol
Download the product protocol from Thermo Fisher Scientific for GeneArt™ Site-Directed Mutagenesis System below.
Download manufacturer protocol
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