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 Constructs
Site-directed mutagenesis of the ERG binding site (−116) within hCLDN5 promoter (CLDN5-mut) was generated using a QuikChange site-directed mutagenesis kit (Stratagene), following the manufacturer's instructions. In brief, pGL3-hCLDN5-WT was used as the template using primers of hCLDN5-ERG-m-F and hCLDN5-ERG-m-R, which resulted in mutated ERG binding element from TTCCT to TTTTT. Primer sequences are listed in supplemental Table 3.

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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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