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What is a three-way ligation - (Nov/05/2005 )

can anybody explain, what is a three-way ligation? what is it advantage?



A three-way ligation is one in which the vector and two pieces of DNA are ligated together in one step. So, you might cut your vector with enzyme X, your first piece of DNA has an enzyme X site on its 5' end and a site for enzyme Y on its 3' end, and your second piece of insert DNA has these sites reversed -- enzyme Y on its 5' end and enzyme X on its 3' end.

Putting it all together, you get:

...---vector--X--1st insert--Y--2nd insert--X--vector--...

where the ...'s are meant to indicate a closed circle...