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

Mutation of Akt Sites in Clk1
Generation of Clk1 mutant cDNA was generated using QuickChange Site-Directed Mutagenesis Kit (Stratagene). The Clk1 template was denatured at 95°C. The mutagenic primers containing the desired mutation(s) were annealed at 55°C and primers extended using PfuUltra DNA polymerase at 68°C. The parental DNA was digested with Dpn I enzyme. The pure mutated DNA was transformed into competent cells and harvested. The mutated fragment was confirmed by 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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