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

The D299N mutant Flag-Reptin cDNA was a gift from M
Cole (Lebanon, USA). The E300G mutant was constructed with the QuikChange site-directed mutagenesis kit from Agilent Technologies (Massy, France) and verified by sequencing. Mutants and WT Flag-Reptin were inserted in the previously described lentiviral vector pRRLSIN-MNDMCS-WPRE (8). Titers were determined by the transduction of HEK 293T cells through the serial dilution of the lentiviral supernatant and were analyzed for p24 protein expression 5 days later.

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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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