The yield of this design was low (<10% on AFM; 48% on TEM, n = 74) and TEM analysis revealed that the internal arrangement of even well-formed bundles is quite disorganized. The majority of malformed structures were categorized as such because of a splitting defect. This was also the case for the rectangle, but that structure still featured a better assembly yield despite containing more folds where splitting could theoretically occur. Additionally, a minority of malformed structures were partial bundles of the expected length but insufficient height or width. These are likely the result of phenomenon known as scaffold fragmentation in DNA origami, wherein a scaffold breaks into two or more shorter fragments. Circular scaffolds can alleviate fragmentation compared to linearized scaffolds, so the twelve-fold bundle was also assembled with the full 60R21N-9kbp plasmid (Supplementary Fig. ). Plasmidfolded structures remain attached to a several kbp vector backbone and this makes it difficult to ascertain their exact yield by AFM. Well-formed bundles were observed in yields that seem at least similar to the purified linearized scaffold, indicating that plasmid-based folding works even though plasmids cannot relieve any torsional strain induced by triplex formation. A further benefit to plasmid-based folding is that it does not require enzymatic cleavage and subsequent purification of the scaffold, thereby avoiding a 70-90% loss of starting material.