Scaffolded DNA origami are a robust tool for building discrete nanoscale objects at high yield. This strategy ensures, in the design process, that the desired nanostructure is the minimum free energy state for the designed set of DNA sequences. Despite aiming for the minimum free energy structure, the folding process which leads to that conformation is difficult to characterize, although it has been the subject of much research. In order to shed light on the molecular folding pathways, this study intentionally frustrates the folding process of these systems by simultaneously annealing the staple pools for multiple target or parent origami structures, forcing competition. A surprising result of these competitive, simultaneous anneals is the formation of chimeric DNA origami which inherit structural regions from both parent origami. By comparing the regions inherited from the parent origami, relative stability of substructures were compared. This allowed examination of the folding process with typical characterization techniques and materials. Anneal curves were then used as a means to rapidly generate a phase diagram of anticipated behavior as a function of staple excess and parent staple ratio. This initial study shows that competitive anneals provide an exciting way to create diverse new nanostructures and may be used to examine the relative stability of various structural motifs.The commercial development of inexpensive and quickly produced DNA of arbitrary nucleobase sequence has fueled the growth of DNA nanotechnology as a field [1]. DNA nanotechnology has made promising advances toward light harvesting [2], computation [3], cancer treatement [4,5], and assembly of nanoelectronics [6]. While many DNA self-assembly strategies exist [7,8], scaffolded DNA origami has received significant attention as a convenient way to design and create discrete nanoscale objects [9]. Scaffolded DNA origami consist of single strand DNA (ssDNA) of two types, synthetic oligomers and circular viral genomes; the viral scaffold is forced to route through the designed structure by the binding of complementary subsequences on the synthetic oligomers, or staples. Staples are added in excess, often 10× relative to the scaffold concentration, strongly driving the viral ssDNA scaffold to fold into the target structure. Although 10× is a common staple excess anneals have been successfully performed as low as 2.5× staple excess. As this study involves multiple staple pools, staple excess will always refer to the total staple concentration relative to the scaffold, while the parent staple ratio will refer to how much of that total corresponds to the staple pool for each target origami. High production yield is achieved by thermal annealing and slow cooling of the system. Such annealing has also been performed chemically, isothermally, and mechanically [10][11][12].The hybridization of double strand DNA (dsDNA) has been well studied, particularly under physiological conditions [13,14]. The formation of dsDNA from ssDNA is driven by base stack...