QuikChange II XL 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

The wild-type human FGF1 coding region was subcloned into a constitutive mammalian expression vector (pcDNA3.1/V5-His, Invitrogen, Waltham, MA, USA) allowing the fusion with a C-terminal V5-His tag (pcDNA3.1-FGF1WT). Mutant FGF1 expression vectors were generated using the QuickChange II XL Site-Directed Mutagenesis Kit (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s protocol. We used the pcDNA3.1-FGF1WT vector as a DNA template to generate pcDNA3.1-FGF1S130A, pcDNA3.1-FGF1S130D, pcDNA3.1-FGF1K132E with specific primers. These vectors allow the expression of C-terminal V5-His tagged wild-type or mutant FGF1 (aa 15 to 154).

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Manufacturer protocol

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

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