QuikChange Site-Directed Mutagenesis Kit, 10 Rxn

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- XL10-Gold cells are resistant to tetracycline and chloramphenicol. If the mutagenized plasmid contains only the tetR or camR resistance marker, an alternative strain of competent cells must be used.	- Use only the Dpn I enzyme provided; do not substitute with an enzyme from another source.

Upstream tips
- XL10-Gold cells are resistant to tetracycline and chloramphenicol. If the mutagenized plasmid contains only the tetR or camR resistance marker, an alternative strain of competent cells must be used.
Protocol tips
- Use only the Dpn I enzyme provided; do not substitute with an enzyme from another source.

Publication protocol

Site-directed mutagenesis
The human K8 cDNA in vector pcDNA3.1 was mutated to generate single-point lysine to arginine or glutamine mutations using the QuikChange site-directed mutagenesis kit (Agilent Technologies). Generation of the truncated mutant of K8 was performed as described previously (). In brief, a frame-shift mutation at Ile465 (ATC→ATCC) generates a truncated 468–amino acid protein (instead of 482). The WT and mutant keratin constructs were confirmed by DNA sequencing.

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Manufacturer protocol

Download the product protocol from Agilent Technologies for QuikChange Site-Directed Mutagenesis Kit, 10 Rxn below.

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