After the plasmids have been digested with the restriction enzymes BamHI and HindIII, you will use the enzyme ligase to make new, or recombinant, DNA.

After you mix the pKAN and pAMP fragments with ligase, you will have many different ligated products. Any BamHI sticky end is capable of binding with any other BamHI sticky end. The same goes for two HindIII sticky ends. However, a BamHI sticky end can not bind to a HindIII sticky end (review the recognition sites for the two enzymes to prove this to yourself).

Below is an animation which illustrates some of the ligation products you might have. The animation starts with an image of the ligation microfuge tube; to run the animation again, reload this page by clicking the "Reload" or "Refresh" button in your browser.