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. 2009 Mar 20;37(9):3061–3073. doi: 10.1093/nar/gkp182

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© 2009 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Ligation-based DNA assembly using restriction endonucleases. (a) DNA assembly using conventional type II endonucleases requires a type II recognition site at the junction between the two starting pieces of DNA. The junction remains in the product, precluding the construction of arbitrary DNA sequences. (b) DNA assembly using type IIS endonucleases, which allow the generation of overhangs of any arbitrary sequence, and thus sequence-independent construction.