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. 2017 Feb 23;45(11):e94. doi: 10.1093/nar/gkx132

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 1. — TPA design and workflow. (A) TPA workflow showing a five-fragment assembly. For step 1, only two of five fragment preparations are represented. After re-annealing, half of the DNA is expected to contain hybridizable overhangs, while the other half is expected to be blunt-end products that will not participate in step 2. For step 2, only one set of compatible fragments with 5΄ overhangs is shown. The set of fragments with 3΄ overhangs should work in a similar fashion. (B) A typical TPA junction showing the long and short primers of two adjacent fragments.