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. 2009 Sep 10;37(20):6984–6990. doi: 10.1093/nar/gkp687

Figure 3.

Figure 3.

Twenty base-pair overlaps are sufficient for oligonucleotide assembly in yeast. (a and b) Schematic demonstrating the assembly of eight 60-mers, named A–H, or their reverse complements, named Arc–Hrc. The oligonucleotides each contain 20 bp overlaps and were assembled into a vector to produce a 340 bp synthetic DNA fragment. The terminal oligonucleotides overlap the vector (grey) by 20 bp (red x). Twenty nucleotide gaps (green) were repaired inside the yeast cell. (c and d) PCR analysis of four randomly selected yeast clones following transformation and assembly of oligonucleotides A–H (c) as depicted in (a) or oligonucleotides A–rc–H–rc (d) as depicted in (b). The predicted amplicon size for a complete assembly is 563 bp and is indicated by an asterisk. M indicates the 100 bp DNA ladder (NEB). (e) Assembly of 28 60-mers, named 1–28 g, containing 20 bp overlaps, to produce a 1140 bp synthetic DNA fragment. (f) PCR analysis of 12 randomly selected yeast clones following transformation and assembly of the oligonucleotides and vector shown in (e). The predicted amplicon size for a complete assembly is 1363 bp and is indicated by an asterisk.