Golden Gate Cloning Pros and Cons
Answer
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What is a key advantage of Golden Gate cloning regarding coding sequences and protein function?
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The key advantage is that the seamless nature of Golden Gate cloning often leaves no restriction enzyme site at the junction of cloned coding sequences. This ensures that the coding sequences remain in frame, resulting in the production of a functional protein.
What are the potential drawbacks related to the length of DNA fragments being cloned using the Golden Gate method? What methods can be used to address unwanted restriction sites in plasmids before Golden Gate cloning?
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As the fragments to be cloned become longer, the probability of an extraneous restriction site interrupting the desired clone increases. This is a potential drawback.
To address unwanted restriction sites in plasmids before Golden Gate cloning, site-directed mutagenesis or PCR can be used.