What criteria do restriction enzymes have to meet when deciding which to choose in order to clone a specific gene into the multiple cloning site of a plasmid (e.g. pUC18)?
The main criteria restriction enzymes must meet for the application in cloning a specific gene into the multiple cloning site of a plasmid are:
- Flank the insert, but does not cut within the insert; - Is in the desired location in the recipient plasmid; - Results in the insert being in the correct orientation in the recipient plasmid; - Is in the frame with tags or fusion proteins in the recipient plasmid.
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