how the multiple cloning site in cloning vector is create? - (Jul/16/2007 )

anybody have any idea how the multiple cloning site in the cloning vector is created?

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design the multiple cloning sites. order primers sense and antisense of the MCS. Anneal them and ligate them into a vector.

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MCS could also be made by adding the sequence to the 5' end of a primer used to amplify a DNA fragment. This DNA fragment along with the multiple cloning site, is then ligated into the vector.

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oppppppsssssss..sorry....maybe i don't make the message right....
regarding your post,my question should be : how to design the multiple cloning site. what i mean is, how the different restriction site for different restriction enzyme is clump together in a particular region, since they not exist in such a way naturally.

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Its like designing a primer. Write down the sequences of the sites you need and order it. Also order the reverse to anneal them.

Now you could add sites so that you can digest the annealed primers and clone it into a vector of choice.

or make a blunt end in the vector and try cloning in the annealed primers.

Good luck !!!

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