We present an analysis of the key steps involved in the DNAdirected assembly of nanoparticles into crystallites and polycrystalline aggregates. Additionally, the rate of crystal growth as a function of increased DNA linker length, solution temperature, and self-complementary versus non-self-complementary DNA linker strands (1-versus 2-component systems) has been studied. The data show that the crystals grow via a 3-step process: an initial ''random binding'' phase resulting in disordered DNA-AuNP aggregates, followed by localized reorganization and subsequent growth of crystalline domain size, where the resulting crystals are well-ordered at all subsequent stages of growth.DNA materials ͉ SAXS ͉ self assembly T he chemical and physical properties of most materials are determined by the placement of individual atoms relative to one another (1-4). These atom-atom interactions include forces such as covalent and ionic bonds, Van der Waals forces and London dispersion forces. However, in the field of nanomaterials, the length scales of particle assembly are significantly larger and the assembly process is governed by a more complicated set of interactions (5-7). The tailorable arrangement of nanoscale materials through directed-mechanisms has proven to be the most practical method to create ordered arrangements of nanoparticles in solution (8-15); crystalline nanoparticle aggregates have potential implications for the development of materials with unique plasmonic (16, 17), photonic (18, 19), electrical (20), and magnetic properties (21). Previous strategies developed to create well-ordered nanoscale assemblies have used electrostatic interactions (11, 12), hydrogen bonding networks (10, 13), and peptide recognition properties (9). Over a decade ago, we introduced the concept of synthetically programmable particle assembly through the use of DNA and polyvalent oligonucleotide nanoparticle conjugates (22). Recently, we (23,24) and the Gang group (25) independently used these principles to construct highly-ordered nanoparticle crystallites via DNA hybridization, where crystal type and lattice parameters can be programmed through design of DNA linker.The formation of these crystals involves multiple types of molecular interactions that are highly dependent and predictable based on the DNA base sequence (22,(26)(27)(28), where the hybridization of the DNA linkers drives the crystallization process. Moreover, the ultimate structure formed is typically the one that maximizes the number of hybridization events, and therefore, if enough thermal energy is provided, the system will typically equilibrate to this structure (23,25,29,30). Indeed, only highly-ordered crystalline aggregates present the thermodynamically most stable arrangement of nanoparticles, because they allow for a maximum of nearestneighbor complementary DNA interactions.Previous work has shown that the complexity of the nanoparticle systems studied thus far necessitates significant thermal annealing to create the thermodynamically favorable crystal structu...