QuikChange Site-Directed Mutagenesis Kit, 10 Rxn

Site Directed Mutagenesis (SDM) Human - Point mutation HeLa Rab11

Experiment
Site Directed Mutagenesis (SDM) Human - Point mutation HeLa Rab11
Product
QuikChange Site-Directed Mutagenesis Kit, 10 Rxn from Agilent Technologies
Manufacturer
Agilent Technologies

Protocol tips

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

In vitro mutagenesis of Rab7 and Rab11 mutant plasmids.
To generate mutant Rab7 and Rab11 plasmids expressing CFP-Rab7Q70L, CFP-Rab7T22N, GFP-Rab11Q70L, and GFP-Rab11S25N (30, 31), we performed in vitro mutagenesis using a QuikChange site-directed mutagenesis kit. The wild-type plasmids CFP-Rab7 and GFP-Rab11 were used as the templates. Mutagenesis was performed using the following primer pairs: for CFP-Rab7Q70L, 5′-ATATGGGACACAGCAGGACTGGAACGGTTCCAGTCTCTC-3′ and 5′-GAGAGACTGGAACCGTTCCAGTCCTGCTGTGTCCCATAT-3′; for CFP-Rab7T22N, 5′-GATTCTGGAGTCGGGAAGAACTCACTCATGAACCAGTAT-3′ and 5′-ATACTGGTTCATGAGTGAGTTCTTCCCGACTCCAGAATC-3′; for GFP-Rab11Q70L, 5′-ATATGGGACACAGCAGGGCTAGAGCGATATCGAGCTATA-3′ and 5′-TATAGCTCGATATCGCTCTAGCCCTGCTGTGTCCCATAT-3′; and for GFP-Rab11S25N, 5′-GATTCTGGTGTTGGAAAGAATAATCTCCTGTCTCGATTT-3′ and 5′-AAATCGAGACAGGAGATTATTCTTTCCAACACCAGAATC-3′. The sequences of the mutant DNA fragments were confirmed by DNA sequencing.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Site Directed Mutagenesis (SDM) Human - Point mutation HeLa Rab11 using QuikChange Site-Directed Mutagenesis Kit, 10 Rxn from Agilent Technologies.

Paper title
Intracellular Transport of Vaccinia Virus in HeLa Cells Requires WASH-VPEF/FAM21-Retromer Complexes and Recycling Molecules Rab11 and Rab22
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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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