QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn

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	All mutations were engineered into the shuttle E vector, using QuikChange II XL site-directed mutagenesis kit (Stratagene) according to the manufacturer’s protocol.

Protocol tips
All mutations were engineered into the shuttle E vector, using QuikChange II XL site-directed mutagenesis kit (Stratagene) according to the manufacturer’s protocol.

Publication protocol

DENV2 full-length cDNA clones with NS5 E111A, E111Q and E111R mutations were constructed with a pACYCNGC FL and a TA-NGC (shuttle E) vector as previously described (1). The pACYC-NGC FL plasmid contains a long DNA fragment comprising a T7 promoter, the DENV2 NGC genome, and the hepatitis delta virus ribozyme sequence (HDVr). The shuttle vector contains nucleotides 5427 to 10955 (from NS3 to 3’UTR and HDVr sequence) from the pACYC-NGC FL plasmid. All mutations were engineered into the shuttle E vector, using QuikChange II XL site-directed mutagenesis kit (Stratagene) according to the manufacturer’s protocol. The primers used for site directed mutagenesis are listed in Table S3A. The mutants were cloned into pACYC-NGC FL at BspEI and MluI restriction sites. All constructs were verified by DNA sequencing before proceeding to the subsequent experiments.

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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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