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. Author manuscript; available in PMC: 2011 Jul 20.
Published in final edited form as: Nat Biotechnol. 2010 Nov 28;28(12):1295–1299. doi: 10.1038/nbt.1716

Figure 2. Gene synthesis products.

Figure 2

GFPmut3 was PCR assembled (a) from two different assembly subpools (GFP42 and GFP35) that were amplified from OLS Pool 1. Because the majority of the products were of the wrong size, we gel-purified the full-length assemblies and re-amplified them (b). Using the longer oligonucleotides in OLS Pool 2 we were able to develop a PCR assembly protocol that did not require gel-isolation, which we used to build three different fluorescent proteins (c). We then attempted to build 42 scFv regions that contained challenging GC-rich linkers. Of the 42 assemblies (d) 40 resulted in strong bands of the correct size. We gel isolated and re-amplified the two that did not assemble (7 and 24) resulting in bands of the correct size (see Supplementary Fig. 10b online). The antibody that corresponds to each number is given in Supplementary Table 3 online.