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. 2014 Sep 16;9(9):e107329. doi: 10.1371/journal.pone.0107329

Figure 3. Cycled Ligation Enables Both Complex and Multiplexed Assemblies.

Figure 3

(A) Complex assembly of very small (80 bp) and large (2100 bp) DNA fragments ( Table 1 ) into a single product. Six DNA fragments of various sizes were seamlessly assembled in a single reaction and amplified by PCR. (B) Sixteen 100–200 bp DNA sequences ( Table 1 ) were placed in a single cycled ligation reaction. Twelve SOCs were added to guide the assembly of four different products consisting of four inserts each (three SOCs per product). Each of the four products was PCR amplified from the single cycled ligation reaction, demonstrating that multiplexed assembly reactions work well. (C) Assembly of sixteen distinct DNA fragments into a single ordered product in a single reaction. The number of inserts (n) assembled into each product is displayed above lanes. All sequences, primers, and SOCs described in Tables S1 and S2.