Hello, I am relatively inexperienced with certain molecular biology techniques. I have a gene that has been cloned into a plasmid for me. I would like to leave the N and C Terminal domains intact but I would like to remove a stretch of twenty amino acids from the center. What would be the most efficient way to approach this? Does anyone have any good protocols I can look at? Thank you.
Site-directed deletion is the most simple and efficient way to achieve that, the following link is the published method (http://bmcbiotechnol.biomedcentral.com/articles/10.1186/1472-6750-8-91) and the attached file is my lab protocol based on that publication for site-directed mutation, insertion and deletion. It is efficient and you can get your deletion done just by one PCR amplification.
Would this method be able to delete 20-30 amino acids from the middle of a plasmid?
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