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help in digesting DNA fragment using restriction enzyme - (Oct/08/2011 )


I am trying to creat a research plan or more like designing my experiment.

I want to cut the DNA fragment with my gene of interest. I want to ligate it with a vector. I need the DNA fragment with restriction sites that I can use to digest the vector and then ligate them together. How can i do that? I mean, how can i add restriction sites to the DNA frgament with my gene of interest,



You have to PCR-amplify your gene of interest using primers which contain the restriction site as an overhanging part of the primer. Then you can digest your PCR product and ligate it into the vector.



Thanks a lot!