Removing a restriction site by linearising and religating a plasmid

Being able to remove a restriction site by digesting to linearise a plasmid, Klenow treating then religating the linear fragment is a very useful trick and an often requested feature.

This is now possible with the new Cloning Clipboard in MacVector 12.7.

  • select the enzyme recognition site you want to remove
  • Click DIGEST to linearize the plasmid and places it on the Cloning Clipboard.
  • Switch to the Cloning Clipboard and view your new fragment
  • Drag one end of the fragment to the other end
  • Modify the ends to blunt them both
  • A new circular sequence will be created from your digested fragment.

    The origin of your plasmid will have changed to the digested site, so you may want to rotate the sequence to show the original sequence.

    RemovingRE Site

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