text: Concatenation of the tags, as developed in 2004, is different from that seen in SAGE. The cleavage of the tags with Xho1 and mixture of the different samples, followed by ligation, form the first concatenation step. The second step uses one of the restriction endonucleases with consensus to the adapter molecule attached to the 3’ end. They are again ligated, and PCR is performed to purify samples for the next joining. The concatenation is continued with the second restriction endonuclease,  followed by the third and finally the fourth. This results in the concatamer formed by the six endonuclease ligations containing 32 tags, arranged 5’ to 5’ around the Xho1 site. In SAGE, concatenation takes place after ditags are formed and amplified by PCR. The linkers on the outside of the ditags are cleaved with the enzyme that provided their binding and these sticky end ditags are concatenated randomly and placed into a cloning vector.